Novel 3-acyloxymethyl-cephem compounds useful as intermediates for preparing cephalosporin antibiotics

ABSTRACT

Novel 3-acyloxymethyl-cephem compounds of the formula: ##STR1## wherein R 1  is hydrogen or an acyl group; W is acetonyl, or a group represented by --X--COOH or --X--OH (X is an organic residue) or salts thereof were found to be useful as starting materials for preparing cephalosporins of the formula: ##STR2## wherein R 3  stands for a residue of a nucleophilic compound and R 1  has the same meaning as above.

This is a continuation of application Ser. No 882,914, filed Mar. 2,1978, now abandoned, which in turn is a continuation of application Ser.No. 660,408, filed Feb. 23, 1976, now abandoned.

The present invention relates to novel 3-acyloxymethyl-cephem compoundsand preparations thereof. More particularly, this invention relates tothe compounds of the formula; ##STR3## wherein R¹ is hydrogen or an acylgroup; W is acetonyl or a group represented by --X--COOH or --X--OH (Xis an organic residue) or salts thereof, for example pharmaceuticallyacceptable salts thereof, and also relates to processes for producingthem.

Cephalosporin derivatives with a 3-hydroxymethyl moiety were onlyobtainable by enzymatic cleavage of the 3-acetyl group from3-acetoxymethyl-cephalosporins or by separating them from thecephalosporin C fermentation byproduct. Recently, it has become possibleto produce7-(D-5-amino-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-carboxylicacid (desacetylcephalosporin C, DCPC) in high titer by fermentation(U.S. Pat. No. 3,926,726, Nature New Biology, 246, 154(1963)) and,alongside of cephalosporin C, this substance has been attractingattention as a starting material for cephalosporin compounds that couldbe more potent in antibiotic activity.

It has, however, been believed to be difficult to acylate the3-hydroxymethyl group of the 3-hydroxymethyl compound (cephalosporadesicacid). For example, Heyningen [Van Heyningen: J.Med.Chem., 8, 22(1965),Advan. Drug. Res., 4, 28(1968)] reported that the O-acylation ofcephalosporadesic acid was feasible only with the use of a large excessof aroyl chloride (yield 32%-57%) and that the use of ketene, aliphaticacid chloride or acetic anhydride did not cause the O-acylation, orinduced a lactonization. Kukolja [J. Med. Chem. 13, 1114(1970)] reporteda roundabout process for the synthesis of O-acyloxymethylcephalosporinswhich comprised O-acylating a 3-hydroxymethyl-2-cephem compound and thencausing the latter to isomerize to the 3-cephem compound. U.S. Pat. No.3,532,694 and Japanese Patent Publication No. 33080/1975 disclosed aprocess in which, to prevent the lactonization reaction, the 4-carboxylgroup of cephalosporadesic acid is first protected, for example byesterification and, then, the O-acylation is carried out. Disclosed inJapanese Patent Application Laid Out No. 42792/1972 is a process whichcomprises O-acylating cephalosporadesic acid with azolide. However,these processes are not commercially profitable because they provideonly low yields or/and involve troublesome and time-consuming proceduresor/and expensive reagents, for instance. Thus, for example, theesterification reaction of cephalosporadesic acid cannot be accomplishedby an ordinary esterification process in which the rearrangement of thedouble bond or the lactonization predominates. While it is possible tointroduce such limited groups as methyl, ethyl, diphenylmethyl, benzyl,etc. by means of diazo compounds such as diazomethane, diazoethane,diphenyldiazomethane, phenyldiazomethane, etc., it is difficult, after3-acylation, to de-esterify the compound without accompaniment of someside reaction such as a fission of the β-lactam ring or a shift of thedouble bond.

On the other hand, the reaction by which the 3-acetoxymethyl group of acephalosporin compound is substituted with a nucleophilic reagententails a concurrent decomposition of the starting material,intermediate and product in its course and a protracted reaction time,and therefore results in lower yields [A. B. Taylor, J. Chem. Soc.,7020(1965)]. Thus, it has been desired to have available a derivativepossessing a group which will lend itself more readily to substitutionthan the acetoxy group.

To overcome the foregoing problems we undertook an extensive research,which led us to the finding that the use of diketene or compounds (III)or (IV) hereinafter described as an acylating agent would enablecephalosporadesic acid to be O-acylated in high yield and that theO-acylated cephalosporin thus synthesized would undergo the desiredsubstitution with a nucleophilic compound with great ease. Thisinvention has been developed on the basis of the above findings.

The above compounds (I) include the compounds wherein R¹ is hydrogen,phenylacetyl, phenoxyacetyl, 5-amino-5-carboxyvaleryl whose amino or/andcarboxyl groups may optionally be protected, or any of the groups foundin the 6- or 7-positions of penicillin or cephalosporin derivatives asthe case may be. Thus, for example, the acyl group R¹ may be selectedfrom among aliphatic acyl groups such as formyl, acetyl, propionyl,butyryl, isobutyryl, valeryl, isovaleryl, cyclopentylcarbonyl,cyclohexylcarbonyl, cycloheptylcarbonyl, cyclopentylacetyl,cyclohexadienylacetyl, etc.; aromatic acyl groups such as benzoyl,p-nitrobenzoyl, toluoyl, naphthoyl, etc.; mono-substituted aliphaticacyl groups such as 2-thienylacetyl, cyanoacetyl, acetoacetyl,4-chloro-3-oxobutyryl, 4-bromo-3-oxobutyryl, chloroacetyl, bromoacetyl,4-methylthio-3-oxobutyryl, 4-carbamoylmethylthio-3-oxobutyryl,α-phenoxypropionyl, α-phenoxybutyryl, tetrazolylthioacetyl,tetrazolylacetyl, p-nitrophenylacetyl, trifluoromethylthioacetyl,trifluoromethylsulfinylacetyl, trifluoromethylsulfonylacetyl,cyanomethylthioacetyl, thiadiazolylthioacetyl, p-nitrophenylacetyl,(2-pyridyloxy)acetyl, (2-oxo-4-thiazolin-4-yl)acetyl,(2-imino-4-thiazolin-4-yl)acetyl, (2-thioxo-4-thiazolin-4-yl)acetyl,4-pyridylthioacetyl, (3-sydnone)acetyl, 1-pyrazolylacetyl,2-furylacetyl, (2-oxo-3-methylpyridazinyl(thioacetyl,(2-aminomethylphenyl)acetyl, (2-aminomethylcyclohexenyl)acetyl, etc.;di-substituted aliphatic acyl groups such as α-carboxyphenylacetyl,mandelyl, α-sulfophenylacetyl, α-sulfo-(p-aminophenyl)acetyl,phenylglycyl, (4-hydroxyphenyl)glycyl, (4-methylthiophenyl)glycyl,(4-methoxyphenyl)glycyl, (4-methanesulfinylphenyl)glycyl,(3-methanesulfonamidophenyl)glycyl, 1-cyclohexenylglycyl, thienylglycyl,furylglycyl, cyclohexadienylglycyl, (3,4-dihydroxyphenyl)glycyl, etc.;5-methyl-3-phenyl-4-isoxazolylcarbonyl;3-(2,6-dichlorophenyl)-5-methyl-4-isoxazolylcarbonyl; and so forth. Itshould be understood that the above-mentioned groups are onlyillustrative of the acyl groups that are of use for the purposes of thisinvention, but preferable acyl groups may be represented by the formula:##STR4## wherein R⁴ stands for acetyl, halogenoacetyl, phenyl,p-hydroxyphenyl, thienyl, 2-imino-4-thiazolin-4-yl,2-oxo-4-thiazolin-4-yl, tetrazolyl, phenoxy, 3-amino-3-carboxypropyl,etc. and R⁵ stands for hydrogen, sulfo, amino, hydroxy, etc.

It should also be understood that any functional groups, e.g. aminoor/and carboxyl, in such acyl groups may be suitably protected. Thus,among protective groups for said amino groups are aromatic acyl groupssuch as phthaloyl, benzoyl, p-nitrobenzoyl, toluoyl, naphthoyl,p-tert-butylbenzoyl, p-tert-butylbenzenesulfonyl, phenylacetyl,benzenesulfonyl, phenoxyacetyl, toluenesulfonyl, chlorobenzoyl, etc.;aliphatic acyl groups such as acetyl, valeryl, capryl, n-decanoyl,acryloyl, pivaloyl camphorsulfonyl, methanesulfonyl, chloroacetyl, etc.;esterified carboxyl groups such as tert-butoxycarbonyl, ethoxycarbonyl,isobornyloxycarbonyl, phenyloxycarbonyl, trichloroethoxycarbonyl,benzyloxycarbonyl, β-methylsulfonylethoxycarbonyl, etc.; carbamoylgroups such as methylcarbamoyl, phenylcarbamoyl, naphthylcarbamoyl,etc.; the corresponding thiocarbamoyl groups;2-methoxycarbonyl-1-methylvinyl; and so forth. As protective groups forthe carboxyl groups of said acyls R¹ and the 4-carboxyl group of thecephem ring, there may be mentioned methyl, ethyl, tert-butyl,tert-amyl, benzyl, p-nitrobenzyl, p-methoxybenzyl, benzhydryl,1-indanyl, phenacyl, phenyl, p-nitrophenyl, methoxymethyl, ethoxymethyl,benzyloxymethyl, acetoxymethyl, pivaloyloxymethyl,β-methylsulfonylethyl, methylthiomethyl, trityl, β,β,β-trichloroethyl,silyl groups such as trimethylsilyl, dimethylsilyl, etc.; and so forth.These carboxyl groups may also be in the form of inorganic or organicsalts with alkali metals such as lithium, sodium, potassium, magnesium,etc.; alkaline earth metals such as calcium; or various amines such asdicyclohexylamine, triethylamine, tributylamine, di-n-butylamine,di-n-propylamine and so forth.

The organic residue denoted by X in said compound (I) is usually acarbon chain which is able to form a five- or six-membered ring with##STR5## and which may include a double bond or such an atom or atoms asoxygen, nitrogen or sulfur therein, preferably oxygen. Moreover, thecarbon chain may be such that its carbon atoms carry suitablesubstituent groups. Among examples of such substituents on the carbonchain, there may be mentioned carboxyl, halogen, nitro, alkyl (e.g.,methyl, ethyl, propyl, methylene, ethylene), aralkyl (e.g. benzyl,phenethyl, etc.), aryl (e.g. phenyl, tolyl, etc.), and hydroxy ormercapto groups substituted thereby (e.g. methoxy, p-chlorophenylthio,etc.). Where two or more such substituents are present, they may form aring with the carbon chain. Of those compounds (I), the compounds inwhich W is acetonyl can be produced by reacting a compound of thegeneral formula: ##STR6## (wherein R² is hydrogen or an acyl group) withdiketene. The diketene may be used as well in the form of an adduct withacetone, acetophenone or the like. This is an equimolar reaction, whichmeans that the desired reaction can be accomplished by using (II) anddiketene in equimolar proportions. However, to adjust for the possibledecomposition of diketene which would take place in the presence ofwater or alcohol, an excess of diketene may be employed. Normally thisreaction can be successfully conducted by allowing a3-hydroxymethylcephalosporin (II) and diketene to interact in a suitableinert solvent at a temperature of -30° C. to 40° C. As said suitableinert solvent, there may be employed any of such solvents asdichloromethane, chloroform, dichloroethane, dimethylformamide,dimethylacetamide, tetrahydrofuran, acetonitrile, ethyl acetate,acetone, dioxane, ether, etc. and various mixtures of such solvents. Thereaction of diketene with 3-hydroxymethylcephalosporin (II) proceeds ata high rate, but in consideration of the fact that the reaction ratedepends somewhat on the reaction temperature, and to ensure that thereaction will be carried to completion, this reaction is usuallyconducted for a period of 0.5 to 15 hours. If necessary, an amine suchas triethylamine may be added to the reaction system, or where thestarting material (II) is an alkali metal salt, an equivalent of, forexample, triethylamine hydrochloride may be added to effect a saltinterchange before the desired reaction takes place.

Where W stands for --X--COOH, the compound can be produced by reacting a3-hydroxymethylcephalosporin (II) with a compound of the generalformula: ##STR7## (where X has the same meaning as definedhereinbefore).

As specific examples of compound (III), there may be mentioned maleicanhydride, succinic anhydride, phthalic anhydride, glutaric anhydride,diglycolic anhydride, thiodiglycolic anhydride, p-chlorophenylsuccinicanhydride, methylenesuccinic anhydride, 3-nitrophthalic anhydride,trimellitic anhydride, isatoic anhydride.

Generally speaking, the reaction of compound (II) with compound (III) isexpediently conducted in the presence of a suitable inert solvent likethat used in the reaction described just above. The stoichiometricproportions of reactants, reaction temperature and other conditions ofreaction may also be similar to those used for the reaction described.

The compound wherein W is --X--OH may be produced by reacting a3-hydroxymethylcephalosporin (II) with a compound of the generalformula: ##STR8## (wherein X has the same meaning as definedhereinbefore).

Thus, as examples of compound (IV), there may be mentionedO-carboxymandelic anhydride, O-carboxy-α-hydroxypropionic anhydride,O-carboxy-β-hydroxypropionic anhydride, O-carboxy-3-methylsalicylicanhydride, O-carboxy-(α-hydroxy-α-phenyl)propionic anhydride,O-carboxy-(α-hydroxy-β-phenyl)propionic anhydride and so forth.

The conditions of reaction between compound (II) and compound (IV) aresimilar to those used for the reactions previously described. Where thestarting material (II) has an unprotected amino group, the reactionthereof with diketene, compound (III) or compound (IV) may be conductedso that both the 3-hydroxy and the amino group may be simultaneouslyacylated by the same acyl groups.

The resultant compound (I) not only has antibiotic activity as such butis ready to react with a nucleophilic compound to introduce the residueof said nucleophilic compound into the 3-methyl group of thecephalosporin to give the compound represented by the formula: ##STR9##wherein R³ stands for a residue of a nucleophilic compound and R¹ hasthe same meaning as defined before. As to the nucleophilic compound usedfor this reaction, any of the compounds that are able to replace the3-acetoxy groups of cephalosporins may be used. Furthermore, thereaction proceeds at a rate ranging from 4 to 16 times that of the3-acetoxy compound and almost quantitatively.

Therefore, among such nucleophilic compounds are nitrogen-containingheterocyclic thiols which contain one or more nitrogen atoms which mayoptionally be in the form of oxide or/and which contain such atoms asoxygen or/and sulfur in addition to the nitrogen atom, with or withoutnuclear substitution. As the common examples of the nitrogen-containingheterocyclic group of such a thiol compound, there may be mentionedpyridyl, N-oxidepyridyl, pyrimidyl, pyridazinyl, N-oxidepyridazinyl,pyrazolyl, diazolyl, thiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,2,3-oxadiazolyl,1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1H-tetrazolyl, 2H-tetrazolyl and soforth. As the substituents on such nitrogen-containing heterocyclicgroups, there may be mentioned such monovalent groups as hydroxy,mercapto, amino, carboxyl, carbamoyl, lower alkyl (for example, methyl,ethyl, trifluoromethyl, propyl, isopropyl, butyl, isobutyl, etc.), loweralkoxy (for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy,etc.), halogens (e.g. chlorine, bromine, etc.), and various substituentgroups as attached through lower alkylene groups, --S--, --N-- or otherpolyvalent groups. When such polyvalent groups are lower alkylenegroups, the substituents may for example be hydroxy, mercapto, amino,morpholino, carboxyl, sulfo, carbamoyl, alkoxycarbonyl, loweralkylcarbamoyl, alkoxy, alkylthio, alkylsulfonyl, acyloxy,morpholinocarbonyl and so forth. When such a polyvalent group is --S--or --N--, the substituents may be lower alkyls or lower alkylene groupshaving the aforementioned substituents. When the polyvalent group is--N--, such substituents as carboxyl, alkoxycarbonyl, acyl, carbamoyl,lower alkylcarbamoyl, etc. may be directly attached. More particularly,there may be mentioned substituted alkyl groups such as carboxymethyl,carbamoylmethyl, N-lower alkylcarbamoylmethyl (e.g.N,N-dimethylcarbamoylmethyl), hydroxy lower alkyl (e.g. hydroxymethyl,2-hydroxyethyl), acyloxy lower alkyl (e.g. acetoxymethyl,2-acetoxyethyl), alkoxycarbonylmethyl (e.g. methoxycarbonylmethyl,hexyloxycarbonylmethyl, octyloxycarbonylmethyl), methylthiomethyl,methylsulfonylmethyl, N-lower alkylamino lower alkyl (e.g.N,N-dimethylaminomethyl, N,N-dimethylaminoethyl,N,N,N-trimethylammoniumethyl), morpholinomethyl, etc.; substituted aminogroups such as lower alkylamino (e.g. methylamino), sulfo-loweralkylamino (e.g. 2-sulfoethylamino), hydroxylower alkylamino (e.g.hydroxyethylamino), lower alkylamino-lower alkylamino (e.g.2-dimethylaminoethylamino, 2-trimethylammoniumethylamino), acylamino(e.g. acetylamino), 2-dimethylaminoacetylamino,2-trimethylammoniumacetylamino, lower alkoxycarbonylamino (e.g.methoxycarbonylamino), etc.; and substituted thio (mercapto) groups suchas methylthio, 2-hydroxyethylthio, 2-acyloxyethylthio (e.g.2-acetoxyethylthio, 2-phenylacetoxyethylthio, 2-caproyloxyethylthio),carboxymethylthio, alkoxycarbonylmethylthio (e.g. methoxycarbonylthio,hexyloxycarbonylmethylthio), carbamoylmethylthio, N-loweralkylcarbamoylmethylthio (e.g. N,N-dimethylcarbamoylmethylthio),acetylmethylthio, N-lower alkylamino-lower alkylthio (e.g.2-N,N-dimethylaminoethylthio, 2-N,N,N-trimethylammoniumethylthio),morpholinocarbonylmethylthio, 2-sulfoethylthio and so forth. Moreparticularly, there may be mentioned various heterocyclic thiols such astetrazolethiol, methyltetrazolethiol, phenyltetrazolethiol,(2-N,N-dimethylaminoethyl)tetrazolethiol, methylthiadiazolethiol,hydroxyethylthiothiadiazolethiol, methylthiothiadiazolethiol,thiadiazolethiol, carbamoylaminothiadiazolethiol,carbamoylmethylthiothiadiazolethiol, thiazolethiol, methylthiazolethiol,carboxymethylthiazolethiol, triazolethiol, dimethyltriazolethiol,pyrazolethiol, ethoxycarbonylmethyltriazolethiol, imidazolethiol,methyloxadiazolethiol, pyridinethiol, pyrimidinethiol,methylpyridazinethiol, triazinethiol and so forth. In addition, use mayalso be made of such nitrogen-containing heterocyclic compounds asaliphatic or aromatic thiols, e.g. methanethiol, ethanethiol,thiophenol, etc.; thiourea and its derivatives such as N-methylthiourea,N-methyl-N'-pyridylthiourea, etc.; thioamide derivatives such asthiosemicarbazide, thioacetamide, thiobenzamide, etc.; sodiumthiosulfate, sodium sulfite, potassium thiocyanate, sodium azide, etc.;pyridine and pyridine derivatives such as quinoline, picoline, nicotinicacid, nicotinamide, isonicotinamide, isonicotinic acid hydrazide,m-bromopyridine, pyridinesulfonic acid, pyridine-m-carbinol(3-hydroxymethylpyridine), pyridinaldehyde, quinoline, isoquinoline,etc.; and such other nitrogen-containing heterocyclic compounds aspyrazine, pyrazinamide (2-carbamoylpyrazine), pyridazine, pyrimidine,imidazole, 1-methylimidazole, pyrazol and so forth. It is also possibleto employ carbon nucleophilic agents toward which the 3-position isknown to be refractory. As examples of such carbon nucleophilicreagents, there may be mentioned cyanides, pyrrole, substituted pyrrole,indole, acetylene, active methylene compounds, e.g. acetylacetone,acetoacetic acid esters, malonic acid esters, cyclohexane-1,3-dione,triacetylmethane and enamine compounds. Alcohols such as methanol,ethanol, propanol, etc. may also be employed in this reaction.

The substitution reaction between such a nucleophilic compound and acompound of formula (I) is normally conducted in a solvent. While thecommonest solvent is water, the hydrophilic organic solvents inert tothe reaction, e.g. acetone, tetrahydrofuran, dimethylformamide,methanol, ethanol, dimethylsulfoxide, etc., and aqueous solvents such asmixtures of water and such reaction-inert, polar solvents as mentionedabove, are employed with preference.

While compound (I) may be a free compound, it is more advantageous tosubject (I) to the reaction in the form of an alkali metal salt, e.g.the sodium, potassium or other salt, or an organic amine salt, e.g. thetriethylamine, trimethylamine or other amine salt. The nucleophilicagent is also reacted in its free form or as an alkali metal, organicamine or other salt.

The proportion of the nucleophilic compound to be used in this reactionis preferably one equivalent or more with respect to compound (I). Whilethe optimum pH depends upon the particular nucleophilic compound andcompound (I), the reaction is generally carried out under weakly acid toweakly alkaline conditions. The reaction temperature is preferablysomewhere between 40° C. and 70° C., although there are no particularlimits. The reaction time cannot be specified in general terms, either,for it depends upon the reaction temperature, pH, type of nucleophilicreagent and other factors. Roughly speaking, however, the reaction goesto completion in 30 minutes to 2 hours when the reaction temperature is60° C. The reaction may also be carried out in the presence of, as addedto the reaction system, an inorganic salt, e.g. the chloride, bromide,iodide, thiocyanide or nitrate of lithium, sodium, potassium, ammoniumor the like.

The compound (I) wherein R¹ is an acyl group can be converted to thecorresponding compound wherein R¹ is hydrogen by cleaving the 7-acylgroup off in a manner conventional per se (e.g. any of the proceduresset forth in Japanese Patent Publication No. 13862/1966 and No.40899/1970, Japanese Patent Application Laid Open No. 34387/1972, No.95292/1975 and No. 96591/1975, Japanese Patent Publication No.35079/1975, U.S. Pat. No. 3,632,578, etc.).

Into this compound, where R¹ is hydrogen, may be introduced a compoundwhich has previously been mentioned by way of example for R¹ as the 6-or 7-substituents of penicillin or cephalosporin compounds, bypreviously activating the same in a known manner. Thus, for example, thecompound wherein R¹ is 4-halogeno-3-oxobutyryl can be obtained byreacting the compound where R¹ is hydrogen with a4-halogeno-3-oxobutyryl halide which, in turn, may be reacted withthiourea to produce the7-[2-(2-imino-4-thiazolin-4-yl)acetamido]-compound. While it dependssomewhat on the type of 3-substituent, these compounds invariablydisplay excellent antibiotic activity. For example, the compound with1-methyltetrazol-5-ylthiomethyl in 3-position is particularly useful,approximately the same effect being accomplished with this compound at adose about one-fifth that of oephazolin.

The present invention is illustrated in further detail below withreference to examples, but it is to be understood that the examples aresolely for the purpose of illustration and not to be construed aslimitations of the invention, and that many variations may be resortedto without departing from the spirit and scope of the invention. In thisspecification, "g", "mg", "ml", "cm", "Hz", "DMSO", and "decomp." areabbreviations of "gram", "milligram", "milliliter", "centimeter","Herz", "dimethylsulfoxide", and "decomposed", respectively. Resinsnamed "Amberlite" are products manufactured by Rohm & Haas Co. in U.S.A."Celite" and "Sephadex" are marketed by Johns-Manville Sales Corp. andPharmacia A.B., respectively. All the temperatures are uncorrected andthe percentages are all on the weight basis except as specificallydefined. The NMR spectra given therein were measured using a VarianModel HA 100 (100 MHz) or T60 (60 MHz) spectrometer withtetramethylsilane as the internal reference and all δ values are in ppm.The symbol s stands for a singlet, d a doublet, t a triplet, q aquartet, m a multiplet, and J a coupling constant.

EXAMPLE 1

(1) In dichloromethane (50 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.05 g), followed by the addition of diketene(0.92 g). The reaction was carried out at room temperature for 3 hours.After the reaction had been completed, the solvent was distilled offunder reduced pressure and the residue was diluted with water andadjusted to pH 6.0 with sodium hydrogen carbonate. The aqueous solutionwas washed twice with ethyl acetate, brought down to pH 2.0 with 4 N-HCland extracted three times with ethyl acetate. The ethyl acetate solutionwas washed with a saturated aqueous solution of sodium chloride, treatedwith magnesium sulfate and filtered. The solvent was then distilled offunder reduced pressure. To the residue was added ether and the resultantpowder was collected by filtration, washed with ether and dried. Theabove procedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (5.48 g) (yield 93.2%).

IR(KBr): 3350, 1775, 1740, 1715, 1640, 1530 cm⁻¹

NMR(δ in d₆ -DMSO): 1.30-2.40(6H,m), 2.17(3H,s), 3.46(2H,ABq,J=18 Hz),3.60(2H,s), 4.72(1H,t), 4.90(2H,ABq,J=12 Hz), 5.01(1H,d,J=5 Hz),5.62(1H,dd,J=5 & 8 Hz), 7.88(4H,s), 8.80(1H,d,J=8 Hz)

(2) In water (50 ml) was dissolved7β-[D-5-phthalimido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (5.88 g), together with 5-mercapto-1-methyl-1H-tetrazole (1.50 g)and sodium hydrogen carbonate (2.10 g). Following the addition of sodiumchloride (15.0 g), the solution was adjusted to pH 5.0 and, then,reacted at 60° C. for 50 minutes. After cooling, a saturated aqueoussolution of sodium chloride (50 ml) was added and the mixture wasadjusted to pH 1.5 with 4 N-HCl. The resultant solid precipitate wasrecovered by filtration, washed with a saturated aqueous solution ofsodium chloride (20 ml) and dissolved in ethyl acetate (100 ml)-water(20 ml). After separation, the ethyl acetate solution was dried and,following the addition of toluene (50 ml), concentrated. The solidprecipitate was recovered by filtration, washed with toluene-ether anddried. The procedure provided7β-[D-5-phthalimido-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid (5.75 g)(yield 95.5%).

IR(KBr): 3325, 1780, 1730, 1715, 1650, 1545 cm⁻¹

NMR(δ in d₆ -DMSO): 1.40-1.76(2H,m), 2.0-2.4(4H,m), 3.64(2H,ABq,J=19Hz), 3.93(3H,s), 4.30(2H,ABq,J=15 Hz), 4.73(1H,t,J=8 Hz), 5.01(1H,d,J=5Hz), 5.62(1H,dd,J=5 & 9 Hz), 7.85(4H,s), 8.80(d,J=9 Hz)

(3) In water (60 ml) was dissolved7β-[D-5-phthalimido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (5.88 g) together with2-(2-hydroxyethylthio)-5-mercapto-1,3,4-thiadiazole (2.90 g) and sodiumbicarbonate (2.20 g). Following the addition of sodium bromide (30 g),the solution was adjusted to pH 5.5 and reacted at 60° C. for 50minutes. After the reaction was completed, the reaction mixture wasdiluted with water (40 ml) and adjusted to pH 5.0. The aqueous solutionwas washed twice with ethyl acetate, brought down to pH 2.0 with 4 N-HCland extracted three times with a mixture of ethyl acetate andtetrahydrofuran (4:1). The organic layer was washed with a saturatedaqueous solution of sodium chloride, dried over magnesium sulfate,filtered and distilled under reduced pressure to remove the solvent. Tothe residue was added ethyl acetate-ether and the resultant powder wasrecovered by filtration, washed with ether and dried. The procedureprovided 7β-[D-5-phthalimido-5-carboxyvaleramido]-3-[2-(2-hydroxyethylthio)-1,3,4-thiadiazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (6.56 g) (yield 96.6%)

IR(KBr): 3325, 1780, 1715, 1645, 1530 cm⁻¹

NMR(d₆ -DMSO):δ1.30-2.40(6H,m), 3.20-3.80(6H,m), 4.27(2H,ABq,J=12 Hz),4.65(1H,t,J=9 Hz), 4.96(1H,d,J=5 Hz), 5.55(1H,dd,J=5 & 8 Hz),7.87(4H,s), 8.70(1H,d,J=8 Hz)

(4) In a mixture of water (50 ml) and tetrahydrofuran (30 ml) wasdissolved7β-[D-5-phthalimido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (5.88 g) together with2-carbamoylmethylthio-5-mercapto-1,3,4-thiadiazole(2.28 g) and sodiumbicarbonate (2.20 g). The solution was adjusted to pH 5.8 and, then,reacted at 60° C. for 70 minutes. After cooling, water (30 ml) was addedand the aqueous solution was adjusted to pH 5.0, washed twice with ethylacetate and brought down to pH 2.0 with 4 N-HCl. It was then extractedthree times with a solvent mixture of ethyl acetate and tetrahydrofuran(2:1) and the organic layer was washed with a saturated aqueous solutionof sodium chloride and dried over magnesium sulfate. The solvent wasdistilled off under reduced pressure and the residue was treated withethyl acetate. The resultant powder was recovered by filtration, washedwith ethyl acetate and dried. The procedure provided7β-[D-5-phthalimide-5-carboxyvaleramido]-3-(2-carbamoylmethylthio-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid (5.83 g) (yield 84.2%)

IR(KBr): 3430, 3340, 1776, 1717, 1680, 1535 cm⁻¹

NMR(d₆ -DMSO): δ 1.30-2.40(6H,m), 3.57(2H,br), 4.40(2H,s),4.32(2H,ABq,J=12 Hz), 4.70(1H,t,J=8 Hz), 5.0(1H,d,J=5 Hz),5.55(1H,dd,J=5 & 8 Hz), 7.20(1H,broad), 7.60(1H,broad), 7.86(4H,s),8.74(1H,d,J=8 Hz)

EXAMPLE 2

(1) In dichloromethane (50 ml) was dissolved7β-[D-5-benzamido-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (6.69 g). At room temperature, triethylamine(1.01 g) and diketene (1.68 g) were added and the reaction was carriedout for 3.0 hours. After the reaction had been completed, the reactionmixture was treated in the same manner as Example 1. The procedureprovided7β-[D-5-benzamido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (5.12 g) (yield 91.3%).

IR(KBr): 3350, 1780, 1735, 1720, 1640, 1530 cm⁻¹

NMR(δ in d₆ -DMSO): 1.45-2.40(6H,m), 2.18(3H,s), 3.50(2H, ABq,J=19 Hz),4.34(1H,m), 4.88(2H,ABq,J=13 Hz), 5.05(1H,d,J=5 Hz), 5.65(1H,dd,J=5 & 9Hz), 7.27-8.0(5H,m), 8.46(1H,d,J=8 Hz), 8.75(1H,d,J=9 Hz)

(2) In water (20 ml) was dissolved7-[D-5-benzamido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (561 mg) together with potassium iodide (400 mg) and pyridine (212mg). The solution was adjusted to pH 6.5 and, then, reacted at 60° C.for 50 minutes. Following the addition of water (10 ml), the aqueoussolution was adjusted to pH 7.0, washed twice with dichloromethane (5.0ml), readjusted to pH 6.0 and concentrated under reduced pressure. Theresidue was subjected to column chromatography on Amberlite XAD-2,followed by elution with water and, then, with a mixture of water andmethanol. The eluate was concentrated and lyophilized. The procedureprovidedN-{7β-[D-5-benzamido-5-carboxyvaleramido]-3-cephem-3-ylmethyl}pyridinium-4-carboxylatemonosodium salt (352 mg).

IR(KBr): 3360, 3250, 1765, 1645, 1630, 1605, 1575, 1530 cm⁻¹

NMR(D₂ O): δ1.50-2.60(6H,m), 3.14(2H,ABq,J=19 Hz), 4.36(1H,m),5.05(1H,d,J=5 Hz), 5.32(2H,ABq,J=15 Hz), 5.60(1H,dd,J=5 Hz),7.0-9.0(10H,m)

(3) In water (6.0 ml) was dissolved7-[D-5-benzamido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (561 mg) together with thiourea (100 mg) and sodium hydrogencarbonate (253 mg). The solution was adjusted to pH 6.5 and reacted at60° C. for 50 minutes. After cooling, the reaction mixture was subjectedto column chromatography on Amberlite XAD-2, followed by elution withwater and water-methanol. The eluate was concentrated and lyophilized.The procedure providedS-{7-[D-5-benzamido-5-carboxyvaleramido]-3-cephem-3-ylmethyl}-thiouronium-4-carboxylatemonosodium salt (464 mg).

IR(KBr): 3350, 3230, 1762, 1645, 1630, 1600, 1580, 1535 cm⁻¹

NMR(D₂ O): δ1.50-2.50(6H,m), 3.20-3.80(3H,m), 4.40(2H,m), 5.05(1H,d,J=5Hz), 5.53(1H,dd,J=5 Hz), 7.30-7.90(5H,m)

EXAMPLE 3

In dichloromethane (50 ml) was dissolved7β-[D-5-(P-toluenesulfonamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.29 g) and, at room temperature,triethylamine (0.51 g) and diketene (1.26 g) were added. The reactionwas carried out for 4.0 hours, after which time the reaction mixture wastreated as in Example 1. The procedure provided7β-[D-5-(P-toluenesulfonamido)-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (5.64 g) (Yield 92.3%).

IR(KBr): 3275, 1780, 1740, 1730, 1715, 1640, 1535 cm⁻¹

NMR(δ in D₆ -DMSO): 1.30-1.80(4H,m), 2.0-2.4(2H,m), 2.16(3H,s),2.33(3H,s), 3.45(2H,ABq,J=19 Hz), 4.91(2H,ABq,J=12 Hz), 5.0(1H,d,J=5Hz), 5.57(1H,dd,J=5 & 8 Hz), 7.40(4H,m), 7.81(1H,d,J=9 Hz),8.64(1H,d,J=8 Hz)

EXAMPLE 4

In dichloromethane (50 ml) was dissolved7β-[D-5-tert-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.35 g) and, at room temperature,triethylamine (1.01 g) and diketene (1.68 g) were added. The reactionwas carried out for 2 hours, after which time the reaction mixture wastreated as in Example 1. The procedure provided7β-[D-5-tert-butylbenzamido)-5-carboxyvaleramido]-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid (5.78 g)(yield 93.8%).

IR(KBr): 3280, 1780, 1740, 1725, 1710, 1640, 1530 cm⁻¹

NMR(δ in d₆ -DMSO): 1.32(9H,s), 1.50-2.40(6H,m), 2.18(3H,s),3.48(2H,br), 3.57(2H,s), 4.35(1H,m), 4.94(2H,Abq,J=13 Hz), 5.06(1H,d,J=5Hz), 5.62(1H,dd,J=5 & 8 Hz), 7.46(2H,d,J=8 Hz), 7.85(2H,d,J=8 Hz),8.35(1H,d,J=8 Hz), 8.78(1H,d,J=8 Hz)

EXAMPLE 5

In dichloromethane (50 ml) was dissolved7β-[D-5-caprylamido-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (6.01 g), followed by the addition oftriethylamine (1.01 g) and diketene (1.68 g). The reaction was carriedout at room temperature for 3 hours, after which time the reactionmixture was treated as in Example 1. The procedure provided7β-[D-5-caprylamido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (4.36 g)(yield 90.3%).

IR(KBr): 3320, 1780, 1745, 1725, 1715, 1645, 1535 cm⁻¹

NMR(δ in d₆ -DMSO): 0.70-2.40(2 1H,m), 2.22(3H,s), 3.53(2H,broad),3.60(2H,s), 4.18(1H,m), 4.91(2H,ABq,J=12 Hz), 5.04(1H,d,J=5 Hz),5.65(1H,dd,J=5 & 8 Hz), 7.87(1H,d,J=8 Hz), 8.70(1H,d,J=8 Hz)

EXAMPLE 6

In dichloromethane (50 ml) was dissolved7β-phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylic acidtriethylamine salt (4.49 g) and, at room temperature, triethylamine(0.51 g) and diketene (1.26 g) were added. The reaction was conductedfor 2 hours, after which time the solvent was distilled off underreduced pressure. The residue was diluted with water and adjusted to pH6.0. This aqueous solution was extracted twice with ethyl acetate,brought down to pH 2.0 with 4 N-HCl and extracted three times with ethylacetate. The ethyl acetate solution was washed with a saturated aqueoussolution of sodium chloride, treated with magnesium sulfate andfiltered. The solvent was distilled off under reduced pressure and theresidue was recrystallized from ethyl acetate-ether. The above procedureprovided7β-phenylacetamido-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylic acid(4.16 g) (yield 96.3%).

IR(KBr): 3270, 1785, 1745, 1715, 1655, 1540 cm⁻¹

NMR(δ in d₆ -DMSO): 2.14(3H,s), 3.52(6H,broad), 4.86(2H,ABq,J=13 Hz),5.00(1H,d,J=5 Hz), 5.63(1H,dd,J=5 & 9 Hz), 7.22(5H,s), 8.93(1H,d,J=9 Hz)

EXAMPLE 7

A mixture of dichloromethane (300 ml), triethylamine (27 ml) anddimethylaniline (100 ml) was previously cooled to 10° C., and7-[D-5-phthalimido-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (50 g) was dissolved therein. To this solution was addeddichlorodimethylsilane (36 ml) whereupon the internal temperatureincreased to 27° C. The mixture was stirred at that temperature for 30minutes, after which time the internal temperature was brought down to-35° C. Then, phosphorus pentachloride (32.4 g) was added. The mixturewas stirred at -25° C. for 40 minutes and, after cooling to -35° C.,thioacetamide (20 g) was added. The mixture was further stirred at-20°--25° C. for 40 minutes and, after cooling to -30° C., methanol (200ml) was gently added in droplets. Then, at the same temperature, sulfurmonochloride (17 ml) was gently added dropwise. The mixture was stirredfor 20 minutes, after which time it was diluted with water (200 ml) andbrought to pH 3.2 with 40% aqueous potassium carbonate solution. Afterstirring for 60 minutes, the resultant crystals were collected byfiltration and rinsed with water and acetone. The crude crystals thusobtained were suspended in 10% hydrochloric acid (230 ml) and stirred at30° C. for 1 hour. The insolubles were filtered off and the filtrate wascooled to 5°-10° C. and brought to pH 3.3 with potassium carbonate. Thefiltrate was stirred for one hour, then the precipitated crystals werecollected by filtration, rinsed with water and acetone, and dried overphosphorus pentoxide. The procedure provided7-amino-3-(1-methyltetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid(17.0 g).

IR(KBr): 1795 cm⁻¹

NMR(δ in D₂ O NaHCO₃): 3.61 & 3.98(ABq,J=18 Hz,2-CH₂), 4.21(s,tetrazole-CH₃), 5.21(d,J=4.5 Hz,6-H), 5.60(d,J=4.5 Hz,7-H)

Chlorine gas (2.8 g) was bubbled through a solution of diketene (3.3 g)in methylene chloride (160 ml) under stirring and cooling to maintainthe internal temperature at -25° to -35° C. for a period of 100 minutes.Then, the mixture was further stirred at the same temperature for 30minutes. Separately,7-amino-3-(1-methyltetrazol-5-yl)-thiomethyl-3-cephem-4-carboxylic acid(10.0 g) and dibutylamine (7.9 g) were dissolved in methylene chloride(60 ml) and the solution was cooled to -10° C. To this solution, theabove reaction mixture was added dropwise under stirring and cooling tomaintain an internal temperature of -10° to -20° C. for 30 minutes. Themixture was further stirred at that temperature for 40 minutes.Thin-layer chromatography of this reaction mixture revealed the presenceof7-(4-chloro-3-oxobutylamido)-3-(1-methyltetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid. Thiourea (4.64 g) was dissolved in this reaction mixture and theinternal temperature was increased to 17°-19° C. The mixture was stirredat this temperature, whereupon crystals separated out. The crystals wererecovered by suction-filtration, washed with methylene chloride (30 ml)and dried. The procedure provided the[2-(2-imino-4-thiazolin-4-yl)acetamido]-3-(1-methyltetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (12.8 g), melting point: 176°-180° C. (decomp.)

IR(KBr): 1762, 1662 cm⁻¹

NMR(δ in d₆ -DMSO): 3.39(s,CH₂ CO), 3.55 & 3.77(ABq,J=18 Hz,2-CH₂),3.90(s,tetrazole 1-CH₃), 4.21 & 4.36(ABq,J=14 Hz,3-CH₂), 5.03(d,J=5Hz,6-H), 5.66(dd,J=9 & 5 Hz,7-H), 6.23(s,thiazolin 5-H), 6.2-7.1(m,##STR10## 8.85(d,J=9 Hz,--CONH--)

EXAMPLE 8

In dichloromethane (30 ml) was dissolved7β-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acidtriethylamine salt (4.55 g), followed by the addition of succinicanhydride (1.50 g). The mixture was stirred at room temperature for 2hours. The dichloromethane was distilled off and, following the additionof 3% aqueous phosphoric acid solution (100 ml), the residue wasextracted with ethyl acetate (150 ml). The ethyl acetate layer waswashed with a saturated aqueous solution of sodium chloride (100 ml×2),dried (over magnesium sulfate) and concentrated under reduced pressure.The residue was treated with ether and the resultant powder wasrecovered by filtration, washed with ether and dried under reducedpressure over phosphorus pentoxide. The procedure provided7β-(2-thienylacetamido)-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid (4.00 g).

IR(KBr): 1782(β-lactam), 1733(--CO₂ H) cm⁻¹

NMR (δ in d₆ -DMSO): 2.50(4H,--CO(CH₂)₂ --), 3.40 & 3.63(2H,ABq,J=18Hz,2-CH₂), 3.75(2H,s,--CH₂ CONH--), 4.71 & 5.07(2H,ABq,J=13Hz,3-CH₂),5.07(1H,d,J=5 Hz,6-H), 5.68(1H,dd,J=5 & 8 Hz,7-H), 6.90 & 7.30(3H,##STR11## 9.10(1H,d,J=8 Hz,--CONH--)

EXAMPLE 9

In dichloromethane (25 ml) was dissolved7β-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acidtriethylamine salt (3.60 g), followed by addition of phthalic anhydride(1.80 g). The mixture was stirred at room temperature for 2 hours, afterwhich time it was treated in a manner similar to that described inExample 8. The procedure provided7β-(2-thienylacetamido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (3.44 g).

IR(KBr): 1777, 1724, 1650(shoulder) cm⁻¹

NMR(δ in d₆ -DMSO): 3.50 & 3.70(2H,ABq,J=18 Hz,2-CH₂), 3.77(2H,s,--CH₂CO--), 4.91 & 5.29(2H,ABq,J═13 Hz,3-CH₂), 5.10(1H,d,J=5 Hz,6-H),5.70(1H,dd,J=5 & 8 Hz,7-H), 6.92 & 7.32(3H, ##STR12## 7.64(4H, ##STR13##9.12(1H, d,J=8Hz,--CONH--)

EXAMPLE 10

In dichloromethane (40 ml) was dissolved7β-mandelamido-3-hydroxymethyl-3-cephem-4-carboxylic acid triethylaminesalt (4.65 g), followed by addition of succinic anhydride (1.50 g). Themixture was stirred at room temperature for 1 hour and a half. After thereaction had been completed, the mixture was treated as in Example 8 toobtain7β-mandelamido-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid (4.45 g).

IR(KBr): 1776, 1737, 1684(shoulder) cm⁻¹

NMR(δ in d₆ -DMSO): 2.48(4H,--CO(CH₂)₂ --), 3.3-3.7(2H,2-CH₂), 4.69 &5.03(2H,ABq,J=13 Hz,3-CH₂), 4.99(1H, ##STR14## 5.06(1H,d,J=5 Hz,6-H),5.68(1H,dd,J=5 & 8 Hz,7-H), 7.35(5H, ##STR15## 9.31(1H,d,J=8Hz,--CONH--)

EXAMPLE 11

In dichloromethane (7 ml) was dissolved7β-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acidtriethylamine salt (0.68 g), followed by addition of glutaric anhydride(0.34 g). The mixture was stirred at room temperature for 2 and a halfhours. The dichloromethane was distilled off and, following the additionof 3% aqueous phosphoric acid (15 ml), the residue was extracted withethyl acetate (25 ml). The ethyl acetate layer was washed with asaturated aqueous solution of sodium chloride (15 ml×2) and dried (overmagnesium sulfate). Then, upon dropwise addition of a 2 N-solution ofsodium 2-ethylhexanoate in isopropyl alcohol (1.50 ml), there wasobtained a powder. This powder was recovered by filtration, washed withethyl acetate-ether and dried over phosphorus pentoxide. The procedureprovided7β-(2-thienylacetamido)-3-(4-carboxybutyryloxy)methyl-3-cephem-4-carboxylicacid disodium salt (0.42 g).

IR(KBr): 1760, 1736(shoulder), 1661, 1609 cm⁻¹

NMR(δ in D₂ O): 1.90(2H,--CH₂ CH₂ CH₂ --), 2.28(4H,--CH₂ CH₂ CH₂ --),3.21 & 3.61(2H,ABq,J=18 Hz,2-CH₂), 3.78(2H,s, ##STR16##4.4-4.9(2H,3-CH₂), 4.98(1H,d,J=5 Hz,6-H), 5.60(1H,d,J=5 Hz,7-H), 6.95 &7.28(3H, ##STR17##

EXAMPLE 12

In dimethylformamide (50 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.35 g), followed by addition of succinicanhydride (1.50 g). The mixture was stirred at room temperature for 30minutes. After the reaction had been completed, 3% aqueous phosphoricacid (250 ml) was added, followed by extraction with ethyl acetate (500ml). The ethyl acetate layer was washed with water (250 ml×2), dried(over magnesium sulfate) and concentrated under reduced pressure. Then,upon addition of ether, there was obtained a powder. This powder wasrecovered by filtration, washed with ether and dried under reducedpressure over phosphorus pentoxide. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid (6.20 g).

IR(KBr): 1779, 1732, 1640 cm⁻¹

NMR(δ in d₆ -DMSO): 1.28(9H,s, ##STR18## 1.73 & 2.24(6H,--(CH₂)₃ --),2.50(4H,--COCH₂ CH₂ CO--), 3.38 & 3.63(2H,ABq,J=18 Hz,2-CH₂),4.37(1H,--CH--NH--), 4.71 & 5.06(2H,ABq,J=13 Hz,3--CH₂), 5.04(1H,d,J=5Hz,6-H), 5.66(1H,dd,J=5 & 8 Hz,7-H), 7.44 & 7.81(4H, ##STR19##8.43(1H,d,J=8 Hz, ##STR20## 8.80(1H,d,J=8 Hz,--CONH--)

EXAMPLE 13

In dimethylformamide (50 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.35 g), followed by addition of phthalicanhydride (1.63 g). The mixture was stirred at room temperature for 30minutes, after which time it was treated as in Example 12. The procedureprovided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (6.70 g).

IR(KBr): 1784, 1726, 1642 cm⁻¹

NMR (δ in d₆ -DMSO): 1.27(9H,s, ##STR21## 1.72 & 2.22(6H,--(CH₂)₃ --),3.46 & 3.71(2H,ABq,J=18 Hz,2-CH₂), 4.37(1H, ##STR22## 4.89 &5.27(2H,ABq,J=13 Hz,3-CH₂), 5.08(1H,d,J=5 Hz,6-H), 5.67(1H,dd,J=5 & 8Hz;7-H), 7.43 & 7.81(4H, ##STR23## 7.62(4H, ##STR24## 8.42(1H,d,J=8Hz,--CHNHCO--), 8.80(1H,d,J=8 Hz,--CONH--)

EXAMPLE 14

In dimethylformamide (50 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.35 g), followed by addition of maleicanhydride (1.49 g). The mixture was stirred at room temperature for 30minutes, after which time it was treated as in Example 12. The procedureprovided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(3-carboxyacryloyloxy)methyl-3-cephem-4-carboxylicacid (6.05 g).

IR(KBr): 1780, 1727, 1640 cm⁻¹

NMR(δ in d₆ -DMSO): 1.32(9H,s, ##STR25## 1.77 & 2.26(6H,--(CH₂)₃ --),3.41 & 3.65(2H,ABq,J=18 Hz, 2-CH₂), 4.37(1H, ##STR26## 4.80 &5.16(2H,ABq,J=13 Hz,3-CH₂), 5.02(1H,d,J=5 Hz,6-H), 5.66(1H,dd,J=5 & 8Hz,7-H), 6.34(2H,--CH═CH--), 7.44 & 7.82(4H, 8.40(1H,d,J=8Hz,--CHNHCO--), 8.80(1H,d,J=8 Hz,--CONH--)

EXAMPLE 15

In dichloromethane (20 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (1.47 g), followed by addition ofp-chlorophenylthiosuccinic anhydride (0.51 g). The mixture was stirredat room temperature for 1 hour, after which time it was treated as inExample 8. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[3-carboxy-3(or2)-(p-chlorophenylthio)propionyloxy]methyl-3-cephem-4-carboxylic acid(1.50 g).

IR(KBr): 1778, 1728, 1636 cm⁻¹

NMR(δ in d₆ -DMSO): 1.28(9H,s, ##STR27## 1.74 & 2.24 (6H,--(CH₂)₃ --),2.73(2H, ##STR28## 3.3-3.8(2H,2-CH₂), 4.03(1H, ##STR29##4.40(1H,--CH--NH--), 4.73 & 5.07(2H,ABq,J=13 Hz,3--CH₂), 5.02(1H,d,J=5Hz,6-H), 5.68(1H,dd,J=5 & 8 Hz,7-H), 7.42(4H, ##STR30## 7.44 & 7.82(4H,##STR31## 8.44(1H,d,J=8 Hz,--CHNHCO--), 8.82(1H,d,J=8 Hz,--CONH--)

EXAMPLE 16

In dichloromethane (50 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.05 g), followed by addition of succinicanhydride (1.50 g). The mixture was stirred at room temperature for 1hour and a half, after which time it was treated as in Example 8. Theprocedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid (5.43 g).

IR(KBr): 1777, 1710, 1644 cm⁻¹

NMR(δ in d₆ -DMSO): 1.52 & 2.17(6H,--(CH₂)₃ --), 2.50(4H,--(CH₂)₂ --),3.3-3.8(2H,2-CH₂), 4.70 & 5.03(2H,ABq,J=13 Hz,3-CH₂), 4.72(1H,t,J=7 Hz,##STR32## 5.01(1H,d,J=5 Hz,6-H), 5.62(1H,dd,J=5 & 8 Hz,7-H), 7.86(4H,s,##STR33## 8.78(1H,d,J=8 Hz,--CONH--)

EXAMPLE 17

In dichloromethane (50 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.05 g), followed by addition of phthalicanhydride (2.22 g). The mixture was stirred at room temperature for 1hour and a half, after which time it was treated as in Example 8. Theprocedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (6.39 g).

IR(KBr): 1772, 1714, 1643 cm⁻¹

NMR(δ in d₆ -DMSO): 1.52 & 2.18(6H,--(CH₂)₃ --), 3.42 & 3.69(2H,ABq,J=18Hz,2-CH₂), 4.74(1H,t,J=8 Hz, ##STR34## 4.89 & 5.28(2H,ABq,13 Hz,3-CH₂),5.06(1H,d,J=5 Hz,6-H), 5.66(1H,dd,J=5 & 8 Hz,7-H), 7.64(4H, ##STR35##7.86(4H,s, ##STR36## 8.81(1H,d,J=8 Hz,--CONH--)

EXAMPLE 18

7β-(2-Thienylacetamido)-3-(3-carboxyacryloyloxy)-methyl-3-cephem-4-carboxylicacid was synthesized as in Example 8.

IR(KBr): 1780, 1725, 1638 cm⁻¹

NMR(d₆ -DMSO): δ3.43 & 3.76(2H,ABq,J=18 Hz,2-CH₂), 3.75 (2H,s,--CH₂CO--), 4.79 & 5.14(2H,ABq,J=13 Hz,3-CH₂), 5.07(1H,d,J=5 Hz,6-H),5.68(1H,dd,J=5 & 8 Hz,7-H), 6.35(2H,s,--CH═CH--), 6.90 & 7.29(3H,##STR37## 9.10(1H,d,J=8 Hz,--CONH--)

EXAMPLE 9

To a mixture of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (2.20 g) and 3-nitrophthalic anhydride (1.16g) was added dichloromethane (15 ml) together with triethylamine (0.42ml). The resultant solution was stirred at room temperature for 1 hourand a half. After the reaction had been completed, the dichloromethanewas distilled off under reduced pressure and 3% aqueous phosphoric acid(120 ml) and ethyl acetate (160 ml) were added to the residue. The ethylacetate layer was washed with water (80 ml×2), dried (over magnesiumsulfate) and concentrated under reduced pressure. Then, upon addition ofether, there was obtained a powder. This powder was recovered byfiltration, washed with ether and dried under reduced pressure overphosphorus pentoxide. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6 (or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (2.10 g).

IR(KBr): 1783, 1735, 1640 cm⁻¹

NMR(δ in d₆ -DMSO): 1.28(9H,s,--C(CH₃)₃), 1.53 & 2.23(6H,--(CH₂)₃ --),3.43 & 3.70(2H,ABq,J=18 Hz,2-CH₂), 4.37(1H, ##STR38## 4.96 &5.41(2H,ABq,J=13 Hz,3-CH₂), 5.07(1H,d,J=5 Hz,6-H), 5.68(1H,dd,J=5 & 8Hz,7-H), 7.42 & 7.80(4H, ##STR39## 7.8-8.4(3H, ##STR40## 8.44(1H,d,J=8Hz, ##STR41## 8.84(1H,d,J=8 Hz,--CONH--)

EXAMPLE 20

To a mixture of7β-(D-5-phthalimido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (2.12 g) and 3-nitrophthalic anhydride (1.16g) was added dichloromethane (15 ml) together with triethylamine (0.42ml). The solution was stirred at room temperature for 1 hour and a half,after which time it was treated as in Example 19. The procedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carboxy-6 (or3)-nitrobenzoyloxy)-methyl-3-cephem-4-carboxylic acid (2.08 g).

IR(KBr): 1775(shoulder), 1718, 1642 cm⁻¹

NMR(δ in d₆ -DMSO): 1.54 & 2.19(6H,--(CH₂)₃ --), 3.40 & 3.68(2H,ABq,J=18Hz,2-CH₂), 4.73(1H,t,J=7 Hz,-CH-), 4.98 & 5.42(2H,ABq,J=13 Hz,3-CH₂),5.06(1H,d,J=5 Hz,6-H), 5.66(1H,dd,J=5 & 8 Hz,7-H), 7.7-8.4(3H, ##STR42##7.86(4H,s, ##STR43## 8.81(1H,d,J=8 Hz,--CONH--)

EXAMPLE 21

To7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (2.20 g) and trimellitic anhydride (1.15 g)were added dichloromethane (15 ml) and triethylamine (0.84 ml) and theresultant solution was stirred at room temperature for 1 hour. After thereaction had been completed, the mixture was treated in the same manneras Example 19. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2,4 (or5)-dicarboxybenzoyloxy)methyl-3-cephem-4-carboxylic acid (2.14 g).

IR(KBr): 1777, 1724, 1636 cm⁻¹

NMR(δ in d₆ -DMSO): 1.29(9H,s,--C(CH₃)₃), 1.54 & 2.25(6H,--(CH₂)₃ --),3.99 & 3.72(2H,ABq,J=18 Hz,2-CH₂), 4.38 ##STR44## 4.95 &5.30(2H,ABq,J=13 Hz,3-CH₂), 5.11(1H,d,J=5 Hz,6-H), 5.70(1H,dd,J=5 & 8Hz,7-H), 7.43 & 7.82(4H, ##STR45## 7.7-8.3(3H, ##STR46## 8.43(1H,d,J=8Hz, ##STR47## 8.84(1H,d,J=8 Hz,--CONH--)

EXAMPLE 22

In dimethylformamide (10 ml) was suspended desacetylcephalosporin C(2.16g) and, under cooling with ice, concentrated hydrochloric acid (0.83 ml)was added. To the resultant solution was added dimethylformamide (10 ml)together with triethylamine (4.20 ml) and 3-nitrophthalic anhydride(3.86 g), and the mixture was stirred at room temperature for 2 hours.Following this reaction, the mixture was diluted with 3% aqueousphosphoric acid (150 ml) and extracted with ethyl acetate (250 ml×2).The ethyl acetate layer was washed with water (200 ml) and a saturatedaqueous solution of sodium chloride (200 ml), dried (over magnesiumsulfate) and concentrated under reduced pressure. Then, upon addition ofether, there was obtained a powder. This powder was recovered byfiltration, washed with ether and dried under reduced pressure overphosphorus pentoxide. The procedure provided 7β-[D-5-(2-carboxy-6 (or3)-nitrobenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (3.77 g).

IR(KBr): 1780(shoulder), 1729, 1638, 1534, 1348 cm⁻¹

NMR(δ in d₆ -DMSO): 1.67 & 2.23(6H,--(CH₂)₃ --), 3.60(2H,2-CH₂),4.39(1H, ##STR48## 4.97 & 5.40(2H,ABq,J=13 Hz,3-CH₂), 5.08(1H,d,J=5Hz,6-H), 5.68(1H,dd,J=5 & 8 Hz,7-H), 7.5-8.5(7H, ##STR49## --CH-NH--),8.78(1H,d,J=8 Hz,--CONH--)

EXAMPLE 23

In dichloromethane (30 ml) was dissolved7β-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acidtriethylamine salt (4.55 g), followed by addition of O-carboxymandelicanhydride (2.38 g). The mixture was stirred at room temperature for 1hour, after which time the dichloromethane was distilled off. To theresidue was added 3% aqueous phosphoric acid (100 ml, followed byextraction with ethyl acetate (150 ml). The ethyl acetate layer waswashed with a saturated aqueous solution of sodium chloride (100 ml×2),dried (over magnesium sulfate) and concentrated under reduced pressure.Then, upon addition of ether, there was obtained a powder. This powderwas recovered by filtration, washed with ether and dried over phosphoruspentoxide under reduced pressure. The procedure provided7β-(2-thienylacetamido)-3-mandelyloxymethyl-3-cephem-4-carboxylic acid(4.00 g).

IR(KBr): 1778, 1742, 1666 cm⁻¹

NMR(δ in d₆ -DMSO): 3.24 & 3.45(2H,ABq,J=18 Hz,2-CH₂), 3.74(2H,s,--CH₂CONH--), 4.76 & 5.06(2H,ABq,J=13 Hz, 3-CH₂), 5.03(1H,d,J=5 Hz,6-H),5.16(1H,s, ##STR50## 5.68(1H,dd,J=5 & 8 Hz,7-H), 6.90 & 7.34(8H,##STR51## 9.09(1H,d,J=8 Hz,--CONH--)

EXAMPLE 24

In dimethylformamide (40 ml) was dissolved sodium7β-mandelamido-3-hydroxymethyl-3-cephem-4-carboxylate (3.86 g), followedby addition of O-carboxymandelic anhydride (2.67 g). The mixture wasstirred at room temperature for 30 minutes, after which 2% H₃ PO₄ (150ml) was added. The mixture was extracted with ethyl acetate (250 ml) andthe ethyl acetate layer was rinsed with water (150 ml×2), dried (overmagnesium sulfate) and concentrated under reduced pressure. Then, uponaddition of ether, there was obtained a powder. This powder wasrecovered by filtration, washed with ether and dried under reducedpressure over phosphorus pentoxide. The procedure provided7β-mandelamido-3-mandelyloxymethyl-3-cephem-4-carboxylic acid (3.68 g).

IR(KBr): 1777, 1745, 1669 cm⁻¹

NMR(δ in d₆ -DMSO): 3.2-3.7(2H,2-CH₂), 4.7-5.2(5H,3-CH₂,6-H & -CH-x2),5.69(1H,7-H), 7.2-7.6(10H, ##STR52## 8.67(1H,--CONH--)

EXAMPLE 25

In dimethylformamide (50 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.35 g), followed by the addition ofO-carboxymandelic anhydride (2.67 g). The mixture was stirred at roomtemperature for 30 minutes, after which time 3% aqueous phosphoric acid(250 ml) was added, followed by extraction with ethyl acetate (500 ml).The ethyl acetate layer was rinsed with water (250 ml×2), dried (overmagnesium sulfate) and concentrated under reduced pressure. Then, uponaddition of ether, there was obtained a powder. This powder wasrecovered by filtration, washed with ether and dried under reducedpressure over phosphorus pentoxide. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (6.30 g).

IR(KBr): 1778, 1736, 1642 cm⁻¹

NMR(δ in d₆ -DMSO): 1.299H,s,--C(CH₃)₃), 1.73 & 2.23(6H,--(CH₂)₃ --),3.2-3.6(2H,2-CH₂), 4.38(1H,--CH--NH--), 4.75 & 5.04(2H,ABq,J=13Hz,3-CH₂), 5.02(1H,d,J=5 Hz,6-H), 5.16(1H,s, ##STR53## 5.66(1H,dd,J=5 &8 Hz,7-H), 7.2-7.5(5H, ##STR54## 7.43 & 7.81(4H, ##STR55## 8.43(1H,d,J=8Hz,--CH--NH--), 8.80(1H,d,J=8 Hz,--CONH--)

EXAMPLE 26

In dichloromethane (50 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt (7.05 g), followed by addition ofO-carboxymandelic anhydride (2.38 g). The mixture was stirred at roomtemperature for 1 hour, after which time it was treated as in Example23. The procedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (6.25 g).

IR(KBr): 1773, 1715, 1647(shoulder)cm⁻¹

NMR(δ in d₆ -DMSO): 1.54 & 2.22(6H,--(CH₂)₃ --), 3.27 & 3.49(2H,ABq,J=18Hz,2-CH₂), 4.73(1H,t,J=7 Hz, ##STR56## 4.74 & 5.03(2H,ABq,J=13Hz,3-CH₂), 4.98(1H,d,J=5 Hz,6-H), 5.16(1H,s, ##STR57## 5.61(1H,dd,J=5 &8 Hz,7-H), 7.2-7.5(5H, ##STR58## 7.86(4H,s, ##STR59## 8.77(1H,d,J=8Hz,--CONH--)

EXAMPLE 27

In dichloromethane (75 ml) was suspended7β-[D-5-phthalimido-5-carboxyvaleramido]-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (6.51 g). Then, at a temperature not exceeding 10° C.,triethylamine (4.20 ml) was added. To the solution thus obtained wasadded N,N-dimethylaniline (10.0 ml) together with dimethyldichlorosilane(4.40 ml). The mixture was stirred at 20°-25° C. for 30 minutes. Themixture was then cooled to -30° C., followed by addition of phosphoruspentachloride (4.20 g). It was then reacted at -25°±2° C. for 30minutes, after which time methanol (25 ml) was added dropwise at atemperature not exceeding -20° C. The mixture was reacted at -15°--10°C. for 20 minutes and, after the addition of water (50 ml), was stirredvigorously for 5 minutes. The reaction mixture was separated and theaqueous layer was taken, washed with ethyl acetate and adjusted to pH3.2 with 40% aqueous potassium carbonate solution. The resultantcrystals were collected by filtration, washed with water, 50% aqueousmethanol and acetone in the order mentioned and dried. The procedureprovided 7β-amino-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (3.43 g).

IR(KBr): 3170, 1798, 1730, 1700, 1615 cm⁻¹

NMR(δ in D₂ O+NaOD): 3.55(2H,ABq,J=18 Hz), 4.6-5.6(4H,m),7.30-7.90(4H,m)

EXAMPLE 28

In dichloromethane (70 ml) was suspended7β-[D-5-phthalimido-5-carboxyvaleramido]-3-(3-oxybutyryloxy)methyl-3-cephem-4-carboxylicacid (5.88 g). Then, at a temperature not exceeding 10° C.,triethylamine (2.80 ml) was added, followed by the addition ofN,N-dimethylaniline (10.0 ml) and dimethyldichlorosilane (3.13 ml). Themixture was stirred at 20°-25° C. for 30 minutes, after which phosphoruspentachloride (4.20 g) was added at -30° C. The mixture was reacted at-25°±2° C. for 30 minutes, after which time methanol (25 ml) was addeddropwise at a temperature not exceeding -20° C. The reaction was furtherallowed to proceed at -15°--10° C. for 20 minutes and, then, water (50ml) was added at -15°--10° C., followed by vigorous stirring for 5minutes. The water layer was taken, washed with dichloromethane andadjusted to pH 3.5 with a 40% aqueous solution of potassium carbonate.The resultant crystals were recovered by filtration, washed with water,50% aqueous methanol and acetone, and finally dried. The procedureprovided 7β-amino-3-(3-oxybutyryloxy)methyl-3-cephem-4-carboxylic acid(2.84 g).

IR(KBr): 3200, 1800, 1745, 1720, 1622 cm⁻¹

NMR(δ in D₂ O+NaOD): 2.27(3H,s), 3.48(2H,ABq,J=18 Hz), 4.6-5.6(4H,m)

EXAMPLE 29

In dimethylformamide (70 ml) was suspended desacetylcephalosporin C(13.7 g), followed by the addition of concentrated sulfuric acid (2.4ml). To the resultant solution was added triethylamine (29 ml), togetherwith N-carboethoxyphthalimide(8.5 g). The mixture was stirred at 30° C.for 50 minutes, followed by the addition of succinic anhydride (3.0 g).The mixture was stirred for 30 minutes, at the end of which time afurther amount (0.6 g) of succinic anhydride was added. The mixture wasfurther stirred for 1 hour and, then, poured in a cold saturated aqueoussolution of sodium chloride (200 ml). The solution was made acidic withphosphoric acid and extracted three times with ethyl acetate. Theextract was extracted back into a solution of sodium hydrogen carbonate(8 g) in water (150 ml). The water layer was adjusted to pH 1.7 withphosphoric acid and extracted with a mixture of tetrahydrofuran anddichloromethane (1:4). The extract was dried over magnesium sulfate,filtered and, with the addition of triethylamine (18 ml), concentratedto dryness. To the residue was added dichloromethane (200 ml) togetherwith triethylamine (6 ml) and dimethylaniline (30 ml). Then, followingthe addition of dimethyldichlorosilane (21 ml), the mixture was stirredfor 30 minutes, after which it was cooled to -30° C. and phosphoruspentachloride (20 g) was added. The mixture was stirred at -30° C. for30 minutes and, following the addition of methanol (63 ml), it wasfurther stirred for 30 minutes. Then, it was diluted with water (120ml), brought to pH 3.0 and allowed to cool. The resultant crystals wererecovered by filtration. The above procedure provided7-amino-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylic acid (5.4g).

IR(KBr): 1802, 1735, 1720 (shoulder) cm⁻¹

EXAMPLE 30

To7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (6.67 g) was added dichloromethane (60 ml) and, under cooling withice, dimethylaniline (3.78 ml) and triethylamine (4.20 ml) were added.To the resultant solution was added dimethyldichlorosilane (3.87 g),followed by stirring at 8°-15° C. for 1 hour. Then, at -30° C.,dimethylaniline (1.26 ml) and phosphorus pentachloride (4.17 g) wereadded. The mixture was stirred at -30°--20° C. for 2 hours. Then, at-45° C., methanol (30 ml) was added dropwise over a period of 10minutes. After the dropwise addition had been completed, the mixture wasstirred at -10°--5° C. for 40 minutes and, then, water (20 ml) was addeddropwise over 5 minutes. Then, the mixture was adjusted to pH 3.3 withconcentrated aqueous ammonia, whereupon a white slurry separated. Afteran hour of standing under ice-cooling, the precipitate was recovered byfiltration, washed with water, methanol and ether in the order mentionedand dried under reduced pressure over phosphorus pentoxide.

The procedure provided7β-amino-3-mandelyloxymethyl-3-cephem-4-carboxylic acid (2.31 g).

IR(KBr): 1800, 1740, 1621 cm⁻¹

EXAMPLE 31

To 7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[2-carboxy-6 (or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (7.26 g) was addeddichloromethane (60 ml) and, under cooling with ice, dimethylaniline(3.78 ml) and triethylamine (4.20 ml) were added. To the resultantsolution was added dimethyldichlorosilane (3.87 g), followed by stirringat 7°-15° C. for 1 hour. The mixture was cooled to -30° C. anddimethylaniline (1.26 ml) and phosphorus pentachloride (4.17 g) wereadded. The mixture was stirred at -30°--20° C. for 2 hours, after whichtime it was cooled to -45° C. and methanol (30 ml) was added dropwiseover a period of 15 minutes. After the dropwise addition had beencompleted, the mixture was stirred at -10°--8° C. for 40 minutes and,then, water (20 ml) was added dropwise over 10 minutes. The mixture wasthen adjusted to pH 3.4 with concentrated aqueous ammonia, whereupon asubstantially white slurry separated. After 45 minutes' standing underice-cooling, the precipitate was recovered by filtration and washed withwater, methanol and ether in the order mentioned. It was then driedunder reduced pressure over phosphorus pentoxide. The procedure provided7β-amino-3-[2-carboxy-6 (or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (2.93 g).

IR(KBr): 1787, 1734, 1614, 1535, 1350 cm⁻¹

EXAMPLE 32

In water (48 ml) was suspended7β-amino-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylic acid (7.33g) and, at 0°-3° C., 2 N-sodium hydroxide (19.5 ml) was added in smallinstallments, care being used not to allow the pH to exceed 8.5. Then,following the addition of sodium hydrogen carbonate (3.65 g), a solutionof D-α-sulfophenylacetyl chloride (5.0 g) in ethyl acetate (8.8 ml) wasadded dropwise at 0°-5° C. over a period of 1 hour. After the dropwiseaddition had been completed, the reaction was carried out at 0°-5° C.for 20 minutes. The reaction mixture was adjusted to pH 5.5 andseparated. The water layer was taken, degassed and, after confirmingthat the pH was within the range of 5.5 to 6.5, ethanol (800 ml) wasadded over a period of 1 hour. Thereafter, the mixture was stirred for30 minutes, followed by cooling to a temperature not exceeding 5° C. Theresultant crystals were recovered by filtration, washed withethanol-water (10:1) and ethanol in the order mentioned and dried. Theprocedure provided7β-(D-α-sulfophenylacetamido)-3-(2-carboxybenzoyloxy)-methyl-3-cephem-4-carboxylicacid trisodium salt (11.6 g).

IR(KBr): 3350, 1768, 1735, 1670, 1610 cm⁻¹

NMR(δ in D₂ O): 3.44(2H,ABq,J=18 Hz), 5.00(2H,ABq,J=13 Hz),5.06(1H,d,J=5 Hz), 5.08(1H,s), 5.67(1H,d,J=5 Hz), 7.3-7.9(m,9H)

EXAMPLE 33

In water (48 ml) was suspended7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (6.09 g)and, at 0°-3° C., 2 N-sodium hydroxide (10.4 ml) was added in smallinstallments, care being used not to allow the pH to exceed 8.5. Then,following the addition of sodium hydrogen carbonate (3.65 g), a solutionof D-α-sulfophenylacetyl chloride (5.0 g) in ethyl acetate (8.8 ml) wasadded dropwise at 0°-5° C. over a period of 1 hour. After the dropwiseaddition had been completed, the reaction was carried out at 0°-5° C.for 20 minutes. Then, the reaction mixture was adjusted to pH 5.5 andseparated. The water layer was taken, degassed and, after confirmingthat the pH was within the range of 5.5 to 6.5, ethanol (800 ml) wasadded to this aqueous solution (about 80 ml). Thereafter, the mixturewas stirred for 30 minutes, at the end of which time it was cooled to atemperature not exceeding 5° C. The resultant crystals were recovered byfiltration, washed with ethanol-water (10:1) and ethanol, and dried. Theprocedure provided7β-(D-α-sulfophenylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid disodium salt (8.8 g).

IR(KBr): 3300, 1763, 1740, 1680, 1610, 1215, 1047 cm⁻¹

NMR(δ in D₂ O): 2.27(3H,s, ##STR60## 3.29(2H,ABq,J=18 Hz,2-CH₂),4.84(2H,ABq,J=13 Hz,3--CH₂), 5.00(1H,d,J=5 Hz,6-H), 5.07(1H,s, ##STR61##5.70(1H,d,J=5 Hz,7-H), 7.25-7.80(5H,m)

EXAMPLE 34

In dimethylformamide (10 ml) was dissolved7β-(D-α-sulfophenylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic aciddisodium salt (940 mg) together with glutaric anhydride (580 mg) andtriethylamine (404 mg). The mixture was stirred at room temperature for2 hours, after which time the dimethylformamide was distilled off. Tothe residue was added a small amount of water and the aqueous solutionwas desalted with Amberlite IR-120(H), adjusted to pH 6.0 with 1N-sodium hydroxide solution and lyophilized. The lyophilizate wasre-dissolved in water and purified by column chromatography on AmberliteXAD-2. The procedure provided7β-(D-α-sulfophenylacetamido)-3-(4-carboxybutyryloxy)methyl-3-cephem-4-carboxylicacid trisodium salt.

IR(KBr): 1760, 1675, 1620 cm⁻¹

NMR(δ in D₂ O): 1.65-2.60(6H,m), 3.41(2H,q,2-CH₂), 4.83(2H,d,3-CH₂),5.0(1H,s, ##STR62## 5.08(1H,d,6-H), 5.75(1H,d,7-H), 7.47(5H,m)

EXAMPLE 35

In dimethylformamide (3 ml) was dissolved7β-(D-α-sulfophenylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic aciddisodium salt (470 mg) together with succinic anhydride (250 mg) andtriethylamine (200 mg). The mixture was stirred at room temperature for2 hours. It was then diluted with a small amount of water and thedimethylformamide was distilled off under reduced pressure. The residuewas dissolved by the addition of water and purified bycolumn-chromatography on Amberlite XAD-2. The procedure provided7β-(D-α-sulfophenylacetamido)-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid trisodium salt.

IR(KBr): 1765, 1685, 1600 cm⁻¹

NMR(δ in D₂ O): 2.62(4H,s,--CO(CH₂)₂ CO--), 3.38(2H,q,2-CH₂),4.95(2H,3-CH₂), 5.10(1H,s, ##STR63## 5.21(1H,d,6-H), 5.91(1H,d,7-H),7.78(5H,m, ##STR64##

EXAMPLE 36

In chloroform (5 ml) was dissolved7β-(D-α-sulfophenylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acidditriethylamine salt (480 mg), followed by the addition of diketene (300mg). The mixture was stirred at room temperature for 2 hours, afterwhich the solvent was distilled off. The residue was diluted with waterand desalted with Amberlite IR-120(H). The desalted solution wasadjusted to pH 5.7 with 1 N-sodium hydroxide solution and lyophilized.Yield 400 mg. The lyophilizate was purified by column chromatography onAmberlite XAD-2. The procedure provided7β-(D-α-sulfophenylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid disodium salt.

In IR and NMR spectra, this product was found in good agreement with theproduct obtained in Example 33.

EXAMPLE 37

In dichloromethane (80 ml) was suspended7β-amino-3-(3-oxybutyryloxy)methyl-3-cephem-4-carboxylic acid (15.7 g).At -10° C., triethylamine (10.1 g) was added. To the resultant solutionwas added a solution of 4-chloro-3-oxobutyryl chloride (1.41 mMol/g)(44.9 g) dropwise at -20°--15° C. over a period of 20 minutes. After thedropwise addition had been completed, the reaction was carried out at atemperature not exceeding -5° C. for 1 hour. After the reaction, thedichloromethane was distilled off and the residue was dissolved by theaddition of tetrahydrofuran (50 ml), ethyl acetate (100 ml) and 10%aqueous phosphoric acid. The resultant solution was separated and theorganic layer was taken. The water layer was extracted with a solventmixture of ethyl acetate-tetrahydrofuran (5:1). The extracts werepooled, washed with a saturated aqueous solution of sodium chloride,dried over magnesium sulfate and filtered. The solvent was distilled offand the residue was treated with ethyl acetate-ether. The resultedpowder was recovered by filtration, washed with ether and dried. Theprocedure provided7β-(4-chloro-3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (20.4 g).

IR(KBr): 3325, 1782, 1740, 1732, 1715, 1685, 1650 cm⁻¹

NMR(δ in d₆ -DMSO): 2.14(3H,s), 3.55(4H,s), 3.54(2H,br), 4.44(2H,s),4.90(2H,ABq,J=13 Hz), 5.05(1H,d,J=5 Hz), 5.54(1H,q,J=5 & 8 Hz),8.96(1H,d,J=8 Hz)

The dichloromethane solution of 4-chloro-3-oxobutyryl chloride used inthis reaction was prepared by dissolving diketene (84.0 g) indichloromethane (420 ml) and introducing chlorine gas (78.1 g) at-30°--35° C. for one hour.

EXAMPLE 38

While a solution of diketene (0.91 g) in dichloromethane (2 ml) wasstirred at -40°--30° C., bromine (1.82 g) was added dropwise.Separately, 7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (2.6 g) and triethylamine (1.7 g) were dissolved in dichloromethane(20 ml) and cooled to -40° C. This solution was added to the abovereaction mixture. After stirring for 20 minutes, the mixture was furtherstirred under cooling with ice. Following the addition of water (7 ml),phosphoric acid and ethyl acetate, the mixture was stirred vigorouslyand the organic layer was washed with an aqueous solution of sodiumchloride, dried and decolorized with activated carbon. Then, the solventwas distilled off and ether was added. The procedure provided7β-(4-bromo-3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (2.4 g).

IR(KBr): 1790, 1730, 1645, 1545 cm⁻¹

NMR(δ in d₆ -DMSO): 2.17(3H,s,COCH₃), 3.60(2H,s,--COCH₂ CO--),3.3-3.8(2H,broad,2--CH₂), 4.36(2H,s,BrCH₂ CO--), 4.76 & 5.06(2H,ABq,J=12Hz,3--CH₂), 5.07(1H,d,J=4.5 Hz,6-H), 5.68(1H,dd,J=4.5 & 8 Hz,7-H),9.04(1H,d,J=8 Hz,--CONH--)

EXAMPLE 39

A solution of diketene (0.20 ml) in dichloromethane (1 ml) was stirredat -30° C., a 1.5 M solution of chlorine in carbon tetrachloride (2.0 g)was added dropwise over a period of 10 minutes. The mixture was furtherstirred at -25°--35° C. for 30 minutes. Separately,7β-amino-3-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (0.90 g) andtriethylamine (0.84 ml) were dissolved in dichloromethane (5 ml) andcooled to -5°--10° C. To this solution was added the above reactionmixture dropwise at -20°--30° C. over a period of 15 minutes. Themixture was then stirred at the same temperature for 45 minutes. Then,the reaction mixture was distilled under reduced pressure and theresidue was stirred vigorously with ethyl acetate (25 ml),tetrahydrofuran (5 ml) and 10% aqueous phosphoric acid (20 ml). Theorganic layer was washed with a saturated aqueous solution of sodiumchloride (15 ml), dried over sodium sulfate and concentrated underreduced pressure. Then, upon addition of ether, there was obtained apowder. This powder was recovered by filtration, washed with ether (10ml) and dried under reduced pressure over phosphorus pentoxide. Theprocedure provided 7β-(4-chloro-3-oxobutylamido)-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (0.70 g).

NMR(δ in d₆ -DMSO): 3.4-3.8(2H,2--CH₂), 3.54(2H,s,--COCH₂ CO--),4.52(2H,s,ClCH₂ --), 4.94 & 5.22(2H,ABq,J=13 Hz,3--CH₂), 5.04(1H,d,J=5Hz,6-H), 5.68(1H,dd,J=5 & 8 Hz,7-H), 7.7-8.5(3H,m, ##STR65##9.04(1H,d,J=8 Hz,--CONH--)

EXAMPLE 40

In acetone (10 ml) was dissolved7β-(4-chloro-3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (4.33 g) and, under cooling with ice, water (10 ml) and thiourea(0.84 g) were added. Then, sodium hydrogen carbonate (0.84 g) and water(10 ml) were further added. The mixture was reacted at room temperaturefor 5 hours, after which time it was cooled with ice. The resultantcrystals were collected by filtration, washed with water, acetone andether and dried. The procedure provided7β-(2-(2-imino-4-thiazolin-4-yl)acetamido)-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid (4.22 g).

IR(KBr): 1775, 1740, 1710, 1661 cm⁻¹

NMR(δ in d₆ -DMSO): 2.17(3H,s, ##STR66## 3.38(2H,s,--CH₂ CONH--),3.51(2H,2--CH₂), 3.59(2H,s ##STR67## 4.77 & 5.06(2H,ABq,J=13 Hz,3--CH₂),5.05(1H,d,J=5 Hz,6-H), 5.69(1H,dd,J=5 & 9 Hz,7-H),6.23(1H,s,thiazoline-H), 8.82(1H,d,J=9 Hz,--CONH--)

EXAMPLE 41

In dimethylformamide (3.5 ml) was dissolved7β-(2-(2-imino-4-thiazolin-4-yl)acetamido)-3-hydroxymethyl-3-cephem-4-carboxylicacid sodium salt (1.37 g), followed by the addition of O-carboxymandelicanhydride (0.90 g). The mixture was stirred at room temperature for onehour, after which time most of the dimethylformamide was distilled offunder reduced pressure. To the residue was added ethyl acetate (50 ml),followed by vigorous stirring. The resultant powder was recovered byfiltration, washed with ethyl acetate (20 ml), dichloromethane (20 ml)and ether (20 ml) in the order mentioned. The procedure provided7β-(2-(2-imino-4-thiazolin-4-yl)acetamido)-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (1.60 g).

IR(KBr): 1780, 1743, 1665, 1643, 1537 cm⁻¹

NMR(δ in d₆ -DMSO+D₂ O): 3.33 & 3.65(2H,ABq,J=18 Hz,2--CH₂),3.37(2H,s,--CH₂ CO--), 4.8-5.3(2H,3--CH₂), 4.97(1H,d,J=5 Hz,6-H),5.21(1H,s, ##STR68## 5.64(1H,d,J=5 Hz,7-H), 6.25(1H,s,thiazolin-H),7.2-7.6(5H, ##STR69##

EXAMPLE 42

In N,N-dimethylformamide (20 ml) was dissolved7β-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acidsodium salt (3.86 g) and, at -5° C., triethylamine (1.40 ml) anddiketene (1.50 ml) were added. The reaction was carried out at -5°-0° C.for 1 hour, after which time the reaction mixture was poured inice-water (200 ml). The mixture was adjusted to pH 2.0 with4N-hydrochloric acid and extracted with ethyl acetate. The ethyl acetatelayer was rinsed with water, diluted with water, brought to pH 7.0 with5% aqueous sodium hydrogen carbonate solution and separated. The waterlayer was taken, concentrated and subjected to column-chromatography onAmberlite XAD-2, elution being carried out with water-methanol. Theeluate was lyophilized. The above procedure provided7β-(2-thienylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid sodium salt (4.48 g).

IR(KBr): 3300, 1765, 1745, 1670, 1613 cm⁻¹

NMR(δ in D₂ O): 2.31(3H,s), 3.47(2H,ABq,J=18 Hz), 3.85(2H,s),4.88(2H,ABq,J=13 Hz), 5.08(1H,d,J=5 Hz), 5.60(1H,d,J=5 Hz,6.9-7.5(3H,m)

EXAMPLE 43

In dichloromethane (50 ml) was dissolved7β-phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylic acidtriethylamine salt (4.5 g), followed by the addition of succinicanhydride (1.5 g). The mixture was stirred at room temperature for 5hours, after which time the solvent was distilled off under reducedpressure. Then, following the addition of water and ethyl acetate, theresidue was adjusted to pH 2.0 with phosphoric acid. The ethyl acetatelayer was dried and concentrated. The resultant crystals were collectedby filtration (2.9 g). The mother fluid was further concentrated and,after the addition of ether, the concentrate was allowed to stand,whereupon crystals (1.6 g) were obtained. These crystals wererecrystallized from ethyl acetate.

The procedure provided7β-phenylacetamido-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid, melting point: 86°-89° C.

IR(KBr): 1800, 1735, 1692, 1660 cm⁻¹

NMR(δ in d₆ -DMSO): 2.48(4H,--(CH₂)₂ --), 3.51(4H,2--CH₂,--CH₂ CO--),4.69 & 5.02(2H,ABq,J=13 Hz), 5.00(1H,d,J=5 Hz,6-H), 5.63(1H,dd,J=5 & 9Hz,7-H), 7.23(5H, ##STR70## 9.02(1H,d,J=9 Hz,--CONH--).

EXAMPLE 44

The reaction procedure of Example 43 was repeated except that phthalicanhydride (2.2 g) was used in place of succinic anhydride. Thisprocedure provided7β-phenylacetamido-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid. Melting point: 128°-129° C. (ethyl acetate-ether)

IR(KBr): 1788, 1731, 1695, 1662 cm⁻¹

NMR(δ in d₆ -DMSO): 3.53(2H,s,--CH₂ CO--), 3.61(2H,2--CH₂), 4.90 &5.27(2H,ABq,J=13 Hz,3--CH₂), 5.08(1H,d,J=5 Hz,6-H), 5.68(1H,dd,J=5 & 8Hz,7-H), 7.25(5H, ##STR71## 7.62(4H, ##STR72## 9.07(1H,d,J=8Hz,--CONH--)

EXAMPLE 45

Dichloromethane (20 ml) was added to a mixture of7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (3.14 g)and dimethylacetamide (1.8 g). Then, under ice-cooling and stirring,phenoxyacetyl chloride (1.8 g) was added. The mixture was stirred for 1hour, after which the insolubles were filtered off and the filtratewashed with an aqueous solution of sodium chloride. The organic layerwas extracted with aqueous sodium hydrogen carbonate solution. The waterlayer was made acidic with phosphoric acid and extracted with ethylacetate. The extract was washed with aqueous sodium chloride solution,dried and distilled to remove the solvent. To the residue was addedether and the resultant powder was recovered by filtration and washedwith ether.

The procedure provided7β-phenoxyacetamido-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid.

IR(KBr): 1788, 1722 cm⁻¹

NMR(δ in d₆ -DMSO): 2.18(3H,s, ##STR73## 3.60(2H,2--CH₂), 3.66(2H,s,##STR74## 4.67(2H,s,--OCH₂ --), 4.99(2H,3--CH₂), 5.18(1H,d,J=5 Hz,6-H),5.78(1H,dd,J=5 & 8 Hz,7-H), 6.8-7.7(5H,m), 9.07(1H,d,J=8 Hz)

EXAMPLE 46

In dichloromethane (5 ml) was suspended7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (314 mg)and, at -10° C, triethylamine (0.28 ml) was added. To the resultantsolution was added diketene (0.2 ml), and the reaction was carried outat a temperature not exceeding 0° C. for 2 hours. After this reaction,the dichloromethane was distilled off and the residue was dissolved inwaterethyl acetate, adjusted to pH 2.0 with 4 N-hydrochloric acid andseparated. The ethyl acetate layer was taken, diluted with water,adjusted to pH 7.0 with 5% aqueous sodium hydrogen carbonate, andseparated. The water layer was taken, concentrated and subjected tocolumn-chromatography on Amberlite XAD-2, elution being carried out withwater-methanol. The eluate was lyophilized. The above procedure provided7β-(3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid sodium salt (375 mg).

IR(KBr): 3320, 1770, 1745, 1660, 1610 cm⁻¹

NMR(δ in D₂ O): 2.27(6H,s), 3.55(2H,ABq,J=18 Hz), 4.93(2H,ABq,J=13 Hz),5.15(1H,d,J=5 Hz), 5.70(1H,d,J=5 Hz)

EXAMPLE 47

In acetonitrile (5 ml) was dissolved7β-(4-bromo-3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (2.4 g), followed by the addition of thiocarbamic acid O-methylester (0.6 g). The mixture was stirred at room temperature overnight andthe resultant crystals were recovered by filtration. The procedureprovided7β-(2-(2-oxo-4-thiazolin-4-yl)acetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid.

IR(KBr): 1780, 1722, 1675, 1629 cm⁻¹

NMR(δ in d₆ -DMSO): 2.17(3H,s,--COCH₃), 3.32(2H,s,--CH₂ --CONH), 3.41 &3.64(2H,ABq,J=18 Hz,2-H), 3.58(2H,s,--COCH₂ CO--), 4.75 &5.06(2H,ABq,J=13 Hz), 5.06(1H,d,J=4.5 Hz,6-H), 5.68(1H,dd,J=4.5 & 8Hz,7-H), 5.99(1H,s,thiazolin-H), 8.94(1H,d,J=8 Hz,--CONH--),11.06(1H,thiazolin-NH)

EXAMPLE 48

In dichloromethane (7 ml) was suspended7β-amino-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylic acid (660mg), followed by the addition of N,N-dimethylacetamide (0.7 ml). Then,under ice-cooling and stirring, a solution of (1H-tetrazol-1-yl)acetylchloride (294 mg) in dichloromethane (2 ml) was added. The mixture wasstirred at room temperature for 1 hour, after which time it was pouredin an aqueous solution of sodium hydrogen carbonate and separated. Thewater layer was taken, washed with dichloromethane, made acidic withphosphoric acid and extracted with ethyl acetate. The extract wasre-extracted with aqueous sodium hydrogen carbonate solution to bringthe desired compound into the aqueous phase. This aqueous solution waspurified by column-chromatography on Sephadex LH-20. The fractionscontaining the dominant product were pooled and lyophilized. Theprocedure provided7β-[2-(1H-tetrazol-1-yl)acetamido]-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid disodium salt.

IR(KBr): 1765, 1620 cm⁻¹

NMR(δ in D₂ O): 2.60(4H,m,--(CH₂)₂ --), 3.44 & 3.74(2H,ABq,J=17Hz,2--CH₂), 5.20(1H,d,J=5 Hz,6-H), 5.59(2H,s,NCH₂ CO--), 5.76(1H,d,J=5Hz,7-H), 9.33(1H,s,tetrazol-H)

EXAMPLE 49

In dichloromethane (10 ml) was suspended7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (942 mg),followed by the addition of N,N-dimethylacetamide (1 ml). Then, underice-cooling and stirring, a solution of (1H-tetrazol-1-yl)acetylchloride (441 mg) in dichloromethane (3 ml) was added. The mixture wasfurther stirred at room temperature for 30 minutes, after which it waspoured in an aqueous solution of sodium hydrogen carbonate. The waterlayer, i.e. aqueous extract, was purified by column-chromatography onSephadex LH-20. The fractions rich in the desired product were pooled,concentrated, made acidic with phosphoric acid and extracted with ethylacetate. The extract was dried, concentrated and treated with ether. Theprocedure provided7β-[2-(1H-tetrazol-1-yl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid.

IR(KBr): 1782, 1707 cm⁻¹

NMR(δ in d₆ -DMSO): 2.17(3H,s,--CH₃), 3.55(2H,broad,2--CH₂), 3.59(2H,s,##STR75## 4.78 & 5.08(2H,ABq,J=13 Hz,3--CH₂), 5.09(1H,d,J=5 Hz,6-H),5.34(2H,s,NCH₂ CO--), 5.71(1H,dd,J=5 & 8 Hz,7-H), 9.28(1H,s,tetrazol-H),9.46(1H,d,J=8 Hz,--CONH--)

EXAMPLE 50

In dry tetrahydrofuran (30 ml) was dissolvedD-α-t-butoxycarbonylamino-α-(p-hydroxyphenyl)acetic acid (2.68 g),followed by the addition of 2,6-lutidine (1.08 g). While cooling at -10°C. and stirring, ethyl chloroformate (1.08 g) was gently added and themixture was stirred at -10° C. for 20 minutes. An ice-cooled mixedsolution of 7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (3.14 g) and sodium hydrogen carbonate (1.0 g) in water (30 ml) wasadded in a single dose to the above mixture. The entire mixture wasstirred at -3° C. for 10 minutes and, then, under cooling withice-water, for 2 hours. Following the addition of water (60 ml), themixture was washed with ethyl acetate (50 ml) and, under stirring in thepresence of ethyl acetate (100 ml), 50% phosphoric acid was gently addedso as to bring the pH to 3.0. The ethyl acetate layer was taken, rinsedwith water (100 ml), dried over anhydrous sodium sulfate and treatedwith a 2 N-solution of sodium 2-ethylhexanoate in isopropyl alcohol (6ml). The resultant precipitate was taken by decantation, loosened withethyl acetate, recovered by filtration and dried under reduced pressureover phosphorus pentoxide. The brown powder thus obtained was purifiedby columnchromatography on Amberlite XAD-2. The procedure provided7β-[D-α-t-butoxycarbonylamino-α-(p-hydroxyphenyl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid sodium salt.

IR(KBr): 3400, 1770, 1680, 1610 cm⁻¹

NMR(δ in D₂ O): 1.45(9H,s), 2.30(3H,s), 3.43(2H,broad), 5.65(1H,d,J=5Hz,7-H), 6.76-7.40(4H,m)

EXAMPLE 51

In water (0.3 ml) was dissolved7β-(D-α-sulfophenylacetamide)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid disodium salt (278 mg), together with pyridine (60 mg) and KSCN(1.2g). The mixture was heated at 60° C. for 1 hour, after which it wassubjected to column-chromatography on Amberlite XAD-2, elution beingcarried out with water. The fractions rich in the desired product werepooled, lyophilized and recrystallized from methanol. The procedureprovided7-(D-α-sulfophenylacetamido)-3-cephem-3-pyridiniummethyl-4-carboxylatesodium salt.

IR(KBr): 1760, 1665, 1610 cm⁻¹

NMR(δ in D₂ O). 2.97, 3.35(2H,ABq,J=18 Hz,2--CH₂), 5.27,5.40(2H,3--CH₂), 5.07(1H,d,J=5.2 Hz,6-H), 5.71(1H,d,βJ=5.2 Hz,7-H),5.10(1H,s, ##STR76## 7.47(5H,m), 8.04,8.55,8.90(5H, ##STR77##

EXAMPLE 52

The reaction procedure of Example 51 was repeated using isonicotinamide(90 mg) in place of pyridine and the reaction product was treated in thesame manner as Example 51. The procedure provided7-(D-α-sulfophenylacetamido)-3-cephem-3-(4-carbamoylpyridinium)methyl-4-carboxylatesodium salt. It was purified by recrystallization from ethanol-water.Melting point: 175° C.(decomp.)

IR(KBr): 1765, 1692, 1645, 1615, 1029 cm⁻¹

NMR(δ in D₂ O): 2.99, 3.56(2H,ABq,J=18 Hz,2--CH₂), 5.40,5.51(2H,3--CH₂),5.13(1H,d,J=4.8 Hz,6-H), 5.73(1H,d,J=4.8 Hz,7-H), 5.10(1H,s, ##STR78##7.40(5H,m), 8.31, 9.07(4H)

EXAMPLE 53

In 50% aqueous acetone (8 ml) was dissolved7β-(2-thienylacetamido)-3-mandelyloxymethyl-3-cephem-4-carboxylic acid(0.49 g) together with acetylacetone (0.50 g) and sodium hydrogencarbonate (0.17 g), and the solution was stirred at 60° C. for 1 hour.The reaction mixture was brought to room temperature and most of theacetone was distilled off under reduced pressure. To the residue wasadded 5% aqueous phosphoric acid solution (10 ml), together with ethylacetate (20 ml). The ethyl acetate layer was washed with a saturatedaqueous solution of sodium chloride (20 ml), dried over magnesiumsulfate and concentrated under reduced pressure. Then, upon addition ofether, there was obtained a powder. This powder was suspended in water(3 ml) and dissolved by the addition of sodium hydrogen carbonate. Thesolution was subjected to column-chromatography on Sephadex LH-20,elution being carried out with water. The desired fractions were pooledand lyophilized. The procedure provided7β-(2-thienylacetamido)-3-(2-acetyl-3-oxo)butyl-3-cephem-4-carboxylicacid sodium salt (0.28 g). This product was dissolved in water (10 ml),followed by the addition of 10% aqueous phosphoric acid (2 ml) and ethylacetate (20 ml). The ethyl acetate layer was rinsed with water, driedover magnesium sulfate and concentrated under reduced pressure. Then,upon addition of ether, there was obtained a powder. This powder wasrecovered by filtration, washed with ether and dried under reducedpressure over phosphorus pentoxide. The procedure provided7β-(2-thienylacetamido)-3-(2-acetyl-3-oxo)butyl-3-cephem-4-carboxylicacid (0.23 g).

IR(KBr): 1765, 1718 cm⁻¹

NMR(δ in d₆ -DMSO): 2.13 & 2.18(6H,s,(COCH₃)₂), 2.6-3.1(2H,m,3--CH₂),3.31 & 3.56(2H,ABq,2--CH₂), 3.75(2H,s,--CH₂ CO--), 4.14(1H, ##STR79##5.01(1H,d,J=5 Hz,6-H), 5.58(1H,dd,J=5 & 8 Hz,7-H), 6.92 & 7.30(3H,##STR80## 9.04(1H,d,J=8 Hz,--CONH--)

EXAMPLE 54

In 50% aqueous acetone (40 ml) was dissolved7β-(2-thienylacetamido)-3-[2-carboxy-6(or3)-nitrobenzoyloxy]-methyl-3-cephem-4-carboxylic acid (2.65 g) togetherwith acetylacetone (2.42 g) and sodium hydrogen carbonate (1.22 g). Themixture was stirred at 60° C. for 1 hour, after which time it wastreated by a procedure similar to that described in Example 53. Theprocedure provided7β-(2-thienylacetamido)-3-(2-acetyl-3-oxo)butyl-3-cephem-4-carboxylicacid (1.26 g). In IR and NMR spectra, this product was in good agreementwith the product according to Example 53.

EXAMPLE 55

In 50% aqueous acetone (8 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[2-carboxy-6 (or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (0.73 g) togetherwith acetylacetone (0.50 g) and sodium hydrogen carbonate (0.34 g). Themixture was stirred at 60° C. for 1 hour, after which it was treated bya procedure similar to that described in Example 53.

The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-acetyl-3-oxo)butyl-3-cephem-4-carboxylicacid (0.29 g).

IR(KBr): 1767, 1721, 1655, 1635 cm⁻¹

NMR(δ in d₆ -DMSO): 1.30(9H,s,--C(CH₃)₃), 1.76 & 2.25(6H,--(CH₂)₃ --),2.6-3.1(2H,m,3--CH₂), 3.2-3.6(2H,2--CH₂), 4.13(1H,--CH(COCH₃)₂),4.37(1H, ##STR81## 4.99(1H,d,J=5 Hz,6-H), 5.57(1H,dd,J=5 & 8 Hz,7-H),7.43 & 7.83(4H, ##STR82## 8.40(1H,d,J=8 Hz,--CH--NH--), 8.77(1H,d,J=8Hz,--CONH--)

EXAMPLE 56

In 50 % aqueous acetone (8 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (0.67 g), together with acetylacetone (0.50 g) and sodium hydrogencarbonate (0.25 g). The mixture was stirred at 60° C. for 1.5 hours,after which it was treated by a procedure similar to that described inExample 53. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-acetyl-3-oxo)butyl-3-cephem-4-carboxylicacid (0.33 g). In IR and NMR spectra, this product was in agreement withthe product obtained in Example 55.

EXAMPLE 57

In 50% aqueous acetone (14 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[2-carboxy-6 (or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (0.73 g), pyrrole(0.20 g) and sodium hydrogen carbonate (0.25 g). The mixture was stirredat 60° C. for 1 hour, after which it was brought down to roomtemperature and most of the acetone was distilled off under reducedpressure. Then, 5% aqueous phosphoric acid solution (15 ml) and ethylacetate (30 ml) were added. The ethyl acetate layer was washed with asaturated aqueous solution of sodium chloride (20 ml), dried (overmagnesium sulfate) and concentrated under reduced pressure. Then, uponaddition of ether, there was obtained a powder. This powder wassuspended in water (3 ml) and dissolved by the addition of sodiumhydrogen carbonate (0.17 g). The solution was subjected tocolumn-chromatography on Sephadex LH-20 (250 ml), elution being carriedout with water. The fractions containing the desired product were pooledand lyophilized. The above procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-pyrrolyl)methyl-3-cephem-4-carboxylicacid disodium salt (0.29 g).

IR(KBr): 1760, 1600 cm⁻¹

NMR(δ in D₂ O): 1.23(9H,s,-C(CH₃)₃), 1.86 & 2.42(6H,--(CH₂)₃ --), 2.86 &3.25(2H,ABq,J=18 Hz,2-CH₂), 3.42 & 3.73(2H,ABq,J=15 Hz,3-CH₂), 4.51(1H,##STR83## 4.96(1H,d,J=5 Hz, 6-H), 5.61(1H,d,J=5 Hz,7-H),5.93(1H,pyrrole-3-H), 6.07(1H,pyrrole-4H), 6.77(1H,pyrrole-5-H), 7.38 &7.76(4H, ##STR84## 7.80(1H,pyrrole-1-H).

EXAMPLE 58

In 50% aqueous acetone (14 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[2-carboxy-6(or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (0.73 g), togetherwith N-methylpyrrole (0.24 g) and sodium hydrogen carbonate (0.25 g).The mixture was stirred at 60° C. for 1 hour. After the reaction hadbeen completed, the mixture was treated in the same manner as Example57. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(N-methylpyrrol-2-yl)methyl-3-cephem-4-carboxylicacid disodium salt (0.21 g).

IR(KBr): 1757, 1597 cm⁻¹

NMR(δ in D₂ O): 1.21(9H,s,--C(CH₃)₃), 1.87 & 2.45(6H,--(CH₂)₃ --), 2.79& 3.07(2H,ABq,J=18 Hz,2-CH₂), 3.46(3H,s,N-CH₃), 3.51 & 3.87(2H,ABq,J=15Hz,3-CH₂), 4.51(1H, ##STR85## 4.91(1H,d,J=5 Hz,6-H), 5.58(1H,d,J=5Hz,7-H), 5.86(1H,pyrrole-3-H), 5.99(1H,pyrrole-4-H),6.61(1H,pyrrole-5-H), 7.39 & 7.79(4H, ##STR86##

EXAMPLE 59

In 50% aqueous acetone (12 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (0.64 g), together with indole (0.35 g) and sodium hydrogencarbonate (0.17 g). The mixture was stirred at 60° C. for 1 hour.Following the completion of the reaction, the mixture was treated by aprocedure similar to that described in Example 57. The procedureprovided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-indolyl)methyl-3-cephem-4-carboxylicacid disodium salt (0.24 g).

IR(KBr): 1758, 1702, 1600 cm⁻¹

NMR(δ in D₂ O): 1.68 & 2.26(6H,--(CH₂)₃ --), 2.45 & 2.82(2H,ABq,J=18Hz,2-CH₂), 3.61 & 3.86(2H,ABq,J=15 Hz,3-CH₂), 4.6-4.9(2H, ##STR87## &6-H), 5.46(1H,d,J=5 Hz,7-H), 7.0-7.8(10H, ##STR88##

EXAMPLE 60

In 50% aqueous acetone (14 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-carboxy-6(or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (0.70 g), togetherwith indole (0.35 g) and sodium hydrogen carbonate (0.25 g). The mixturewas stirred at 60° C. for 45 minutes and, after the reaction had beencompleted, it was treated by a procedure similar to that described inExample 57. The procedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-indolyl)methyl-3-cephem-4-carboxylicacid disodium salt (0.22 g). In IR spectrum, this product was found toagree with the product obtained in Example 59.

EXAMPLE 61

In water (7 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[2-carboxy-6(or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (0.73 g), sodiumazide (0.26 g) and sodium hydrogen carbonate (0.25 g). The mixture wasstirred at 60° C. for 40 minutes. The reaction mixture was brought toroom temperature and, following the addition of 10% aqueous phosphoricacid (10 ml), extracted with ethyl acetate (30 ml). The ethyl acetatelayer was washed with a saturated solution of sodium chloride (20 ml),dried over magnesium sulfate and concentrated under reduced pressure.Then, upon addition of ether, there was obtained a powder. This powderwas suspended in water (3 ml) and dissolved by the addition of sodiumhydrogen carbonate (0.17 g). This solution was subjected tocolumn-chromatography on Sephadex LH-20 (250 ml), elution being carriedout with water. The fractions containing the desired product were pooledand lyophilized.

The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-azidomethyl-3-cephem-4-carboxylicacid disodium salt (0.36 g).

IR(KBr): 2100, 1766, 1606 cm⁻¹

NMR(δ in D₂ O): 1.32(9H,s,--C(CH₃)₃), 1.92 & 2.48(6H,--(CH₂)₃ --), 3.05& 3.53(2H,ABq,J=18 Hz,2-CH₂), 4.03 & 4.20(2H,ABq,J=13 Hz,3-CH₂),4.52(1H, ##STR89## 5.08(1H,d,J=5 Hz,6-H), 5.68(1H,d,J=5 Hz,7-H), 7.56 &7.86(4H, ##STR90##

EXAMPLE 62

In phosphate buffer (40 ml) of pH 6.4 was dissolved7-[2-(2-imino-4-thiazolin-4-yl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (908 mg), together with a nitrogen-containing heterocyclic thiol(2.2 mMol) and sodium hydrogen carbonate (336 mg). The solution wasstirred at 60° C. for 1 hour, after which it was concentrated underreduced pressure to about 20 ml. The concentrate was subjected tocolumn-chromatography on Amberlite XAD-2, elution being carried out withwater, 5% ethanol and 10% ethanol in the order mentioned. The fractionscontaining the desired product were pooled and lyophilized to obtain thecorresponding one of the following compounds. The reaction yield figurewas the value determined by liquid chromatography immediately followingthe reaction

(1)7-[2-(2-Imino-4-thiazolin-4-yl)acetamido]-3-(2-carboxymethyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid disodium salt; yield 85%

IR(KBr); 1761 cm⁻¹

NMR(δ in D₂ O): 3.56 & 3.92(2H,ABq,J=18 Hz,2-CH₂), 3.76(2H,s,--CH₂ CO),4.16(2H,s,-CH₂ CO), 4.20 & 4.62(2H,ABq,J=13 Hz,3-CH₂), 5.24(1H,d,J=5Hz,6-H), 5.79(1H,d,J=5 Hz,7-H), 6.65(1H,s,thiazolin-5-H).

(2)7-[2-(2-Imino-4-thiazolin-4-yl)acetamido]-3-(3-hydroxymethyl-4-methyl-1,2,4-triazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid sodium salt; yield 82%.

IR(KBr): 1760 cm⁻¹

NMR(δ in D₂ O): 3.40 & 3.82(2H,ABq,J=18 Hz,2-CH₂), 3.62 (2H,s,--CH₂ CO),3.74(3H,s,--CH₃), 3.72 & 4.34(2H,ABq,J=13 Hz,3-CH₂), 4.82(2H,s,CH₂ OH),5.08(1H,d,J=5 Hz,6-H), 5.64(1H,d,J=5 Hz,7-H), 6.52(1H,s,thiazolin-5-H).

EXAMPLE 63

In 50 ml of water was dissolved 5.61 g of7-[D-5-(benzamido)adipinamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, together with 1.50 g of 5-mercapto-1-methyl-1H-tetrazole and 2.20g of sodium bicarbonate. After the pH was adjusted to 5.2, the reactionwas carried out at 60° C. for 50 minutes. After cooling, 100 ml of asaturated aqueous solution of sodium chloride was added and the pH wasadjusted to pH 1.5 with 4 N-HCl. The solid precipitate was recovered byfiltration, rinsed with 20 ml of a saturated aqueous solution of sodiumchloride and dissolved in 100 ml of ethyl acetate-tetrahydrofuran (2:1)and 20 ml of water. The organic layer was dried and the solvent wasdistilled off under reduced pressure. To the residue was addedether-ethyl acetate and the resultant powder was recovered byfiltration, rinsed with ether and dried. The procedure provided 5.45 g(yield 94.8%) of7-[D-5-benzamido-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 3340, 1783, 1730, 1645, 1535 cm⁻¹

NMR(δ in d₆ -DMSO): 1.50-2.0(4H,m), 2.05-2.45(2H,m), 3.70 (2H,broad),3.93(3H,s,N CH₃), 4.15-4.55(3H,m), 5.10 (1H,d,J=5 Hz,6-H),5.66(1H,dd,J=5 & 9 Hz,7-H), 7.32-7.97(5H,m, ##STR91## 8.43(1H,d,J=8Hz,--CONH--), 8.73(1H,d,J=9 Hz,--CONH--)

EXAMPLE 64

In 50 ml of water was dissolved 6.11 g of7-[D-5-(p-toluenesulfonamido)adipinamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, together with 1.50 g of 5-mercapto-1-methyl-1H-tetrazole and 2.20g of sodium bicarbonate. After the solution was adjusted to pH 5.0, thereaction was conducted at 60° C. for 50 minutes. Following the reaction,the reaction mixture was treated in the same manner as Example 63. Theprocedure provided 5.96 g (yield 95.1%) of7-[D-5-(p-toluenesulfonamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 3275, 1780, 1727, 1635, 1535 cm⁻¹

NMR(δ in d₆ -DMSO): 1.45-1.78(4H,m), 2.0-2.3(2H,m), 2.41 (3H,s,--CH₃),3.71(2H,broad,2-CH₂), 3.95(3H,s,--NCH₃), 4.28(2H,broad), 5.06(1H,d,J=5Hz,6-H), 5.62(1H,dd,J=5.0 & 9.0 Hz,7-H), 7.47(4H,m, ##STR92##7.91(1H,d,J=9.0 Hz), 8.68(1H,d,J=9.0 Hz).

EXAMPLE 65

In 50 ml of water was dissolved 6.17 g of7-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, together with 1.74 g of 5-mercapto-1-methyl-1H-tetrazole and 2.50g of sodium bicarbonate. After the solution was adjusted to pH 5.0, thereaction was conducted at 60° C. for 50 minutes. Following the reaction,the mixture was treated in the same manner as Example 63 to obtain 6.02g (yield 95.3%) of7-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 3350, 1780, 1725, 1643, 1532, 1504 cm⁻¹

NMR(δ in d₆ -DMSO): 1.30(9H,s,--C(CH₃)₃), 1.50-2.45(6H,m,--(CH₂)₃ --),3.64(2H,broad,2-CH₂), 3.93(3H,s,--NCH₃), 4.27(2H,broad,3-CH₂),4.36(1H,m, ##STR93## 5.0(1H,d,J=5 Hz,6-H), 5.62(1H,dd,J=5.0 & 8.0Hz,7-H), 7.35(2H,d,J=8.0 Hz), 7.70(2H,d,J=8 Hz), 8.36(1H,d,J=8.0Hz,--CONH--), 8.76(1H,d,J=8.0 Hz,--CONH--)

EXAMPLE 66

In 50 ml of water was dissolved 4.83 g of7-[D-5-(caprylamido)-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, together with 1.74 g of 5-mercapto-1-methyl-1H-tetrazole and 2.50g of sodium bicarbonate. After the solution was adjusted to pH 5.0 and35.0 g of sodium bromide was added, the reaction was conducted at 60° C.for 45 minutes.

Following this reaction period, the reaction mixture was treated in thesame manner as Example 63. The procedure provided 4.78 g (yield 96.3%)of7-[D-5-(caprylamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 3300, 1775, 1725, 1655, 1640, 1545, 1533 cm⁻¹

NMR(δ in d₆ -DMSO): 0.60-2.40(21H,m), 3.70(2H,broad, 2-CH₂),3.95(3H,s,NCH₃), 4.15(1H,m), 4.27(2H,broad, 3-CH₂), 5.03(1H,d,J=5Hz,6-H), 5.65(1H,dd,J=5.0 & 8.0 Hz,7-H), 7.97(1H,d,J=8.0 Hz,-CONH-),8.76(1H, d,J=8.0 Hz,--CONH--)

EXAMPLE 67

In 50 ml of water was dissolved 4.32 g of7-phenylacetamido-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid,together with 1.50 g of 5-mercapto-1-methyl-1H-tetrazole and 1.68 g ofsodium bicarbonate and the reaction was conducted at 60° C. for 50minutes. After cooling, the reaction mixture was adjusted to pH 5.0 andwashed with ethyl acetate. It was then brought down to pH 2.0 andextracted three times with ethyl acetate. The ethyl acetate solution waswashed with a saturated aqueous solution of sodium chloride, treatedwith magnesium sulfate, filtered and distilled under reduced pressure toremove the solvent. The residue was recrystallized from ethylacetate-ether. The procedure provided 4.29 g (yield 96.2%) of7-phenylacetamido-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 3270, 1785, 1733, 1662, 1628, 1542 cm⁻¹

NMR(δ in d₆ -DMSO): 3.55(2H,s,--CH₂ CO--), 3.60(2H,broad,2-CH₂),3.92(3H,s,NCH₃), 4.26(2H,broad,3-CH₂), 5.00(1H,d,J=5.0 Hz,6-H),5.60(1H,dd,J=5.0 & 8.0 Hz,7-H), 7.23(5H,s, ##STR94## 8.98(1H,d,J=8Hz,--CONH--)

EXAMPLE 68

In 10 ml of water was dissolved 561 mg of7-[D-5-benzamido-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, together with 252 mg of sodium bicarbonate and 212 mg of2-methylthio-5-mercapto-1,3,4-thiadiazole. The reaction was conducted at60° C. for 50 minutes and, after cooling, the reaction mixture waswashed with ethyl acetate and freeze-dried. The resultant solid wasdissolved in a small quantity of methanol and treated with acetone. Thecrystals formed were collected by filtration and rinsed with ether. Theprocedure provided 614 mg of7-[D-5-benzamido-5-carboxyvaleramido]-3-(2-methylthio-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid disodium salt.

IR(KBr): 3400, 1766, 1640, 1600, 1530 cm⁻¹

NMR(δ in D₂ O): 1.40-2.55(6H,m,-(CH₂)₃ -), 2.68(3H,s,SCH₃),3.33(2H,ABq,J=18 Hz,2-CH₂), 4.14(2H,ABq,J=14 Hz,3-CH₂), 4.30(1H,m,##STR95## 5.01(1H,d,J=4.5 Hz,6-H), 5.58(1H,d,J=4.5 Hz,7-H),7.25-7.95(5H,m, ##STR96##

EXAMPLE 69

In 10 ml of water was dissolved 561 mg of7-[D-5-benzamido-5-carboxyvaleramido]-5-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, together with 252 mg of sodium carbonate and 224 mg of5-mercapto-2-ethoxycarbonylmethyl1H-1,3,4-triazole. The reaction wasconducted at 60° C. for 50 minutes and, after cooling, the reactionmixture was treated in the same manner as Example 68. The procedureprovided 642 mg of7-[D-5-benzamido-5-carboxyvaleramido]-3-(2-ethoxycarbonylmethyl-1H-1,3,4-triazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid disodium salt.

IR(KBr): 3400, 3280, 1765, 1745, 1640, 1603, 1535 cm⁻¹

NMR(δ in D₂ O): 1.20(3H,t,J=8.0 Hz,--CH₂ CH₃), 1.50-2.50(6H,m,--(CH₂)₃--), 3.32(2H,ABq,J=19 Hz,2-CH₂), 3.80-4.50(7H,m), 4.95(1H,d,J=4.5Hz,6-H), 5.52(1H,d,J=4.5 Hz,7-H), 7.20-7.90(5H,m, ##STR97##

EXAMPLE 70

In 12 ml of water was dissolved 1.27 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid, togetherwith 0.25 g of 5-mercapto-1-methyl-1H-tetrazole and 0.68 gof sodium hydrogen carbonate, followed by the stirring for one hour anda half at 60° C. The reaction solution was allowed to be cooled at theroom temperature and added 30 ml of 4% aqueous solution of phosphoricacid, followed by extracting with 60 ml of ethyl acetate. The ethylacetate layer was washed with a saturated aqueous solution of sodiumchloride (50 ml×2) and dried over magnesium sulfate, followed bycondensation under reduced pressure. The condensate was added ether andthe resultant powder was recovered by filtration, washed with ether anddried over phosphorous pentoxide under reduced pressure to give 1.02 gof7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

In IR and NMR spectra, this product was found in good agreement with theproduct obtained in Example 65.

EXAMPLE 71

In 12 ml of water were dissolved 1.26 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(3-carboxyacryloxy)methyl-3-cephem-4-carboxylicacid, 0.25 g of 5-mercapto-1-methyl-1H-tetrazole and 0.68 g of sodiumhydrogen carbonate, and the solution was stirred for 2 hours at 60° C.,followed by the treatment in the same manner as Example 70. Theprocedure provided 0.88 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid, of which IR and NMR were found in good agreement with that of theproduct obtained in Example 65.

EXAMPLE 72

The reaction of Example 71 was repeated employing 1.36 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid. The procedure provided 0.97 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid, of which IR and NMR were found in good agreement with that of theproduct obtained in Example 65.

EXAMPLE 73

In 12 ml of water were dissolved 1.55 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[3-carboxy-3(or2)-(p-chlorophenylthio)propionyloxy]methyl-3-cephem-4-carboxylic acid,0.25 g of 5-mercapto-1-methyl-1H-tetrazole and 0.68 g of sodium hydrogencarbonate, and the solution was stirred for one hour and a half at 60°C., followed by the treatment in the same manner as Example 70. Theprocedure provided 0.99 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid, of which IR and NMR were found in good agreement with that of theproduct obtained in Example 65.

EXAMPLE 74

The reaction of Example 73 was repeated employing 1.21 g of7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid. The procedure provided 1.00 g of7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid, of which IR and NMR were found in good agreement with that of theproduct obtained in Example 1 (2).

EXAMPLE 75

The reaction of Example 73 was repeated employing 1.30 g of7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid. The procedure provided 0.94 g of7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid, of which IR and NMR were found in good agreement with that of theproduct obtained in Example 1 (2).

EXAMPLE 76

In water (6 ml) was dissolved 7β-[D-5-(2-carboxy-6-(or3)-nitrobenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (0.78 g) togetherwith 5-mercapto-1-methyl-1H-tetrazole (0.12 g) and sodium hydrogencarbonate (0.42 g). The solution was stirred at 60° C. for 30 minutes,after which time it was treated as in Example 70. The procedure provided7β-[D-5-(2-carboxy-6 (or3)-nitrobenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (0.50 g).

IR(KBr): 1782, 1731, 1645, 1537, 1351 cm⁻¹

NMR(d₆ -DMSO): δ 1.73 & 2.26(6H,-(CH₂)₃ -), 3.69(2H,2-CH₂),3.94(3H,s,>N-CH₃), 4.32(2H,3-CH₂), 4.52(1H, ##STR98## 5.06(1H,d,J=5Hz,6-H), 5.67(1H,dd,J=5 & 8 Hz,7-H), 7.6-8.4(4H, ##STR99## 8.79(1H,d,J=8Hz,--CONH--)

EXAMPLE 77

In water (6 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6(or3)-nitrobenzoyloxy) methyl-3-cephem-4-carboxylic acid (0.73 g) togetherwith 5-mercapto-1-methyl-1H-tetrazole (0.12 g) and sodium hydrogencarbonate (0.34 g). The mixture was stirred at 60° C. for 30 minutes,after which time it was treated as in Example 70. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (0.53 g).

In IR(KBr) and NMR(d₆ -DMSO) spectra, this compound was in goodagreement with the product obtained in Example 65.

EXAMPLE 78

The reaction of Example 76 was repeated employing 0.73 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2,4(or5)-dicarboxybenzoyloxy)methyl-3-cephem-4-carboxylic acid to give 0.52 gof7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)-thiomethyl-3-cephem-4-carboxylicacid. In IR(KBr) and NMR(d₆ -DMSO) spectra, this compound was in goodagreement with the product in Example 65.

EXAMPLE 79

In water (6 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid (0.70 g) togetherwith 5-mercapto-2-methyl-1,3,4-thiadiazole (0.13 g) and sodium hydrogencarbonate (0.34 g). The solution was stirred at 60° C. for 30 minutes,after which time it was treated as in Example 65. The procedure provided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-methyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (0.51 g).

IR(KBr): 1773(sh), 1715, 1648(sh)cm⁻¹

NMR(d₆ -DMSO): δ1.53 & 2.15(6H,--(CH₂)₃ --), 2.67(3H,s,--CH₃), 3.45 &3.72(2H,ABq,J=18 Hz,2-CH₂), 4.19 & 4.50(2H,ABq,J=13 Hz,3-CH₂),4.72(1H,t,J=7 Hz, ##STR100## 5.02(1H,d,J=5 Hz,6-H), 5.61(1H,dd,J=5 & 8Hz,7-H), 7.87(4H,s, ##STR101## 8.72(1H,d,J=8 Hz,--CONH--)

EXAMPLE 80

The reaction of Example 79 was repeated employing 0.73 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid to give 0.55 g of7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(2-methyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 1780, 1728, 1644 cm⁻¹

NMR(d₆ -DMSO): δ 1.28(9H,s,--C(CH₃)₃), 1.74 & 2.23(6H,--(CH₂)₃),2.66(3H,s,--CH₃), 3.50 & 3.75(2H,ABq,J=18 Hz,2-CH₂), 4.20 &4.50(2H,ABq,J=13 Hz,3-CH₂), 4.39(1H, ##STR102## 5.05(1H,d,J=5 Hz,6-H),5.65(1H,dd,J=5 & 8 Hz,7-H), 7.44 & 7.80(4H, ##STR103## 8.42(1H,d,J=8 Hz,##STR104## 8.80(1H,d,J=8 Hz,--CONH--)

EXAMPLE 81

In water (5 ml) was dissolved7β-(D-mandelamido)-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid (0.46 g) together with 5-mercapto-1-methyl-1H-tetrazole (0.12 g)and sodium hydrogen carbonate (0.25 g). The solution was stirred at 60°C. for 1 hour and a half. After cooling in the air, the reaction mixturewas subjected to column chromatography on Amberlite XAD-2, elution beingcarried out with water and, then, with a solvent mixture of water andmethanol. The fractions containing the desired compound are pooled,concentrated and lyophilized. The procedure provided7β-(D-mandelamido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid sodium salt (0.31 g)

IR(KBr): 1761, 1675, 1604 cm⁻¹

NMR(D₂ O): δ 3.25 & 3.68(2H,ABq,J=18 Hz,2-CH₂), 3.95(3H,s, >N-CH₃), 4.02& 4.29(2H,ABq,J=13 Hz,3-CH₂), 4.97 (1H,d,J=5 Hz,6-H), 5.18(1H,s,##STR105## 5.51(1H,d,J=5 Hz,7-H), 7.37(5H,s, ##STR106##

EXAMPLE 82

In water (5 ml) was dissolved7β-(2-thienylacetamido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (0.50 g) together with sodium hydrogen carbonate (0.17 g),potassium iodide (0.40 g) and pyridine (0.21 g). The solution wasadjusted to pH 6.5 and, then, stirred at 60° C. for one hour and a half.After cooling in the air, the reaction mixture was subjected to columnchromatography on Amberlite XAD-2, elution being carried out with waterand, then, with a solvent mixture of water and methanol. The fractionscontaining the desired product were pooled, concentrated andlyophilized.

The procedure provided7β-(2-thienylacetamido)-3-(1-pyridylmethyl)-3-cephem-4-carboxylic acidbetaine (0.23 g).

IR(KBr): 1763, 1698, 1617 cm⁻¹

NMR(D₂ O): δ 3.17 & 3.67(2H,ABq,J=17 Hz,2-CH₂), 3.38(2H,s, --CH₂ CO--),5.19(1H,d,J=5 Hz,6-H), 5.41 & 5.67(2H,ABq, J=14 Hz,3-CH₂), 5.75(1H,d,J=5Hz,7-H), 7.01 & 7.28 (3H, ##STR107## 8.15 & 8.62 & 9.04(5H, ##STR108##

EXAMPLE 83

The following compounds were synthesized by procedures similar to thatdescribed in Example 79.

(1)7β-(D-5-Phthalimido-5-carboxyvaleramido)-3-[2-(2-hydroxyethylthio)-1,3,4-thiadiazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid.

IR(KBr): 3325, 1780, 1715, 1645, 1530 cm⁻¹

NMR(d₆ -DMSO): δ 1.30-2.40(m,6H), 3.20-3.80(m,6H), 4.27 (AB-q, 2H,J=12Hz), 4.65(t,1H,J=9 Hz), 4.96(d,1H,J=5 Hz), 5.55(q,1H,J=5 & 8 Hz),7.87(s,4H), 8.70 (d,1H,J=8 Hz)

(2)7β-(D-5-Phthalimido-5-carboxyvaleramido)-3-(2-carbamoylmethylthio-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid

IR(KBr): 3430, 3340, 1776, 1717, 1680, 1535, cm⁻¹

NMR(d₆ -DMSO): δ 1.30-2.40(m,6H), 3.57(broad;2H), 4.40(s,2H), 4.32(AB-q,2H,J=12 Hz), 4.70(t,1H,J=8.0 Hz), 5.0(d,1H,J=5 Hz), 5.55(q,1H,J=5 & 8Hz), 7.20(br.1H), 7.60(br.1H), 7.86(s,4H), 8.74(d,1H,J=5 Hz)

EXAMPLE 84

7β-(2-Thienylacetamido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid was prepared by the procedure similar to that described in Example81. Yield 0.37 g starting from 0.50 g of7β-(2-thienylacetamido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid.

IR(KBr): 1776, 1734, 1672 cm⁻¹

NMR(d₆ -DMSO): δ 3.56 & 3.78(2H,ABq,J=18 Hz,2-CH₂), 3.73(2H,s,--CH₂CO--), 3.92(3H,s,>N-CH₃), 4.21 & 4.37(2H,ABq,J=ABq,J=13 Hz,3-CH₂),5.05(1H,d,J=5 Hz,6-H), 5.66(1H,dd,J=5 & 8 Hz,7-H), 6.90 & 7.29(3H,##STR109## 9.10(1H,d,J=8 Hz,--CONH--)

EXAMPLE 85

In water (12 ml) was dissolved7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (1.33 g) together with 5-mercapto-1-methyl-1H-tetrazole (0.25 g)and sodium hydrogen carbonate (0.51 g). The mixture was stirred at 60°C. for 30 minutes, after which time it was brought down to roomtemperature. Following the addition of 4% aqueous phosphoric acidsolution (30 ml), the reaction mixture was extracted with ethyl acetate(60 ml). The ethyl acetate layer was washed with a saturated aqueoussolution of sodium chloride (40 ml×2), dried (over magnesium sulfate)and concentrated under reduced pressure. Then, upon addition of ether,there was obtained a powder. This powder was recovered by filtration,washed with ether and dried under reduced pressure over phosphoruspentoxide. The procedure provided7β-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (1.14 g).

In IR(KBr) and NMR(d₆ -DMSO) spectra, this compound was good agreementwith the product in Example 65.

EXAMPLE 86

In water (12 ml) was dissolved7β-(D-5-phthalimido-5-carboxyvaleramido)-3-mandelyloxymethyl-3-cephem-4-carboxylicacid (1.27 g) together with 5-mercapto-1-methyl-1H-tetrazole (0.25 g)and sodium hydrogen carbonate (0.51 g). The solution was stirred at 60°C. for 30 minutes. After the reaction had been completed, the reactionmixture was treated in the same manner as Example 85. The procedureprovided7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (1.12 g).

In IR(KBr) and NMR(d₆ -DMSO) spectra, this compound was good agreementwith the product in Example 1 (3).

EXAMPLE 87

In water (5 ml) was dissolved7β-(D-mandelamido)-3-mandelyloxymethyl-3-cephem-4-carboxylic acid (0.50g) together with 5-mercapto-1-methyl-1H-tetrazole (0.12 g) and sodiumhydrogen carbonate (0.17 g). The solution was stirred at 60° C. for 30minutes. After cooling in the air, the reaction mixture was subjected tocolumn chromatography on Amberlite XAD-2, elution being carried out withwater and a solvent mixture of water and methanol. The fractionscontaining the desired product were pooled, concentrated andlyophilized. The procedure provided7β-(D-mandelamido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid sodium salt (0.34 g).

In IR and NMR spectra, this compound was good agreement with the productin Example 81.

EXAMPLE 88

In water (5 ml) was dissolved7β-(2-thienylacetamido)-3-mandelyloxymethyl-3-cephem-4-carboxylic acid(488 mg) together with sodium hydrogen carbonate (84 mg), potassiumiodide (400 mg) and pyridine (212 mg). The solution was adjusted to pH6.5 and reacted at 60° C. for 45 minutes. After cooling, the reactionmixture was subjected to column chromatography on Amberlite XAD-2,elution being carried out with water and, then, a solvent mixture ofwater and methanol. The fractions containing the desired product werepooled, concentrated and lyophilized. The procedure provided7β-(2-thienylacetamido)-3-(1-pyridylmethyl)-3-cephem-4-carboxylic acidbetaine (250 mg).

In IR and NMR spectra, this compound was good agreement with the productin Example 82.

EXAMPLE 89

In water (4 ml) was dissolved7β-(2-thienylacetamido)-3-mandelyloxymethyl-3-cephem-4-carboxylic acid(0.49 g) together with 5-mercapto-1-methyl-1H-tetrazole (0.12 g) andsodium hydrogen carbonate (0.17 g). The solution was stirred at 60° C.for 30 minutes. After the reaction had been completed, the reactionmixture was treated by a procedure similar to that described in Example85. The procedure provided7β-(2-thienylacetamido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (0.40 g).

In IR and NMR spectra, this compound was good agreement with the productin Example 84.

EXAMPLE 90

The following compounds were synthesized by procedures similar to thosedescribed in Example 85 and 86.

(1) 7β-[D-5-(p-tButylbenzamido)-5-carboxyvaleramido]-3-(2-methyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid. Yield 84%.

In IR and NMR spectra, this compound was good agreement with the productin Example 80.

(2)7β-(D-5-Phthalimido-5-carboxyvaleramido)-3-(2-methyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid. Yield 86%.

In IR and NMR spectra, this compound was good agreement with the productin Example 79.

(3)7β-(D-5-Phthalimido-5-carboxyvaleramido)-3-[2-(2-hydroxyethylthio)-1,3,4-thiadiazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid. Yield 81%.

In IR and NMR spectra, this compound was good agreement with the productin Example 83 (1).

(4)7β-(D-5-Phthalimido-5-carboxyvaleramido)-3-(2-carbamoylmethylthio-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid. Yield 88%.

In IR and NMR spectra, this compound was good agreement with the productobtained in Example 83 (2).

EXAMPLE 91

In water (30 ml) were dissolved7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (3.14 g),sodium hydrogen carbonate (1.84 g) and 5-mercapto-1-methyl-1H-tetrazole(1.4 g) and the solution was adjusted its pH to 5.5, followed bystirring and heating at 60° C. for one hour. After cooling, the reactionsolution was washed with dichloromethane (20 ml) and the aqueous layerwas adjusted to pH 3.3, followed by stirring for one hour underice-cooling. The resultant precipitates were collected by filtration andwashed with water, methanol and acetone in this order, followed bydrying to give7β-amino-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (2.7 g).

IR(KBr): 1795 cm⁻¹

NMR(δ in D₂ O+NaHCO₃): 3.61 & 3.98(2H,ABq,J=18 Hz,2-CH₂), 4.21(s,-NCH₃),5.21(d,J=4.5 Hz,6-H), 5.60(d,J=4.5 Hz,7-H)

EXAMPLE 92

In water (30 ml) were dissolved7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (3.14 g),sodium hydrogen carbonate (0.84 g) and5-mercapto-1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazole(2.60 g) and theresultant solution was stirred for 60 minutes at 55° C. After cooling,acetone (15 ml) was added to the reaction solution and the mixture ispassed through a column of active alumina (10 g). The column was washedwith water-acetone (1:1) (30 ml) and the washing was combined with theeluate, followed by distilling off acetone under reduced pressure. Tothe residual solution was added Amberlite IR-120 (acid form) (6.0 ml)and the mixture was stirred for 30 minutes under ice-cooling. Theinsolubles were filtered off and the filtrate was condensed. Thecondensate was added dropwise into ethanol about 30 times volume of thecondensate and precipitated solid was collected by filtration, followedby washing with ethanol and drying to give7β-amino-3-1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazol-5-yl-thiomethyl-3-cephem-4-carboxylicacid (3.28 g).

IR(KBr): 3450, 1780, 1620, 1540 cm⁻¹

NMR(δ in D₂ O): 3.07(6H,s), 3.70(2H,ABq,J=17 Hz), 3.85(2H,t,J=6 Hz),4.25(2H,ABq,J=12 Hz), 4.8-5.2(4H,m)

EXAMPLE 93

In water (30 ml) was dissolved7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (3.14 g)together-with sodium hydrogen carbonate (1.84 g) and2-mercapto-5-methyl-1,3,4-thiadiazole (1.6 g) and the resultant solutionwas adjusted its pH to 6.4, followed by stirring for one hour at 60° C.After cooling, the reaction solution was washed with dichloromethane andpH of the aqueous layer was adjusted to 3.5 under ice-cooling, followedby stirring for one hour. The precipitated materials were collected byfiltration and washed with water, methanol and acetone in this order,followed by drying to give7β-amino-3-(2-methyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid (2.9 g).

IR(KBr): 1795 cm⁻¹

NMR(δ D₂ O+NaHCO₃): 2.87(3H,s,thiadiazole --CH₃), 3.53 &3.95(2H,ABq,J=18 Hz,2-CH₂), 4.10 & 4.46(2H,ABq,J=13 Hz,3-CH₂),5.17(1H,d,J=4.5 Hz,6-H), 5.58(1H,d,J=4.5 Hz,7-H)

EXAMPLE 94

In water (1 ml) containing sodium salt of 5-mercapto-1H-1,2,3-triazole(120 mg) and sodium hydroxide (40 mg) was dissolved under ice-cooling7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid (282 mg)and to the resultant solution was added 1 N-HCl to adjust its pH to 5.5under stirring, followed by further stirring for one hour at 55° C. Tothe reaction solution was added methanol (5 ml) and the mixture wasallowed to cool to the room temperature. The cooled mixture is adjustedits pH to 3.9 by adding 1 N-HCl under stirring and the resultant mixturewas further stirred for one hour under ice-cooling. The precipitatedinsolubles were collected by filtration and washed with water andmethanol in this order. The insolubles were dried naturally and thenover phosphorus pentoxide to give7β-amino-3-(1H-1,2,3-triazol-5-yl)-thiomethyl-3-cephem-4-carboxylic acid(190 mg).

IR(KBr): 1800, 1525 cm⁻¹

EXAMPLE 95

In water (1 ml) containing sodium hydrogen carbonate (84 mg) wasdissolved 7β-amino-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid(314 mg) and to the resultant solution were added isonicotinamide (185mg) and potassium iodide (1.0 g), followed by stirring for one hour at55° C. To the reaction mixture was added ethanol (20 ml) under stirringand resulting precipitates were collected by filtration, followed bywashing with ethanol. After natural drying, the brown power wasdissolved in water (3 ml) and the solution was chromatographed on columnpacked with Amberlite XAD-2. The eluate was freeze-dried to give7β-amino-3-(4-carbamoylpyridinium)methyl-4-carboxylate (150 mg).

IR(KBr): 3500, 1760, 1600 cm⁻¹

EXAMPLE 96

In a mixture of tetrahydrofuran (5 ml) and water (10 ml) were dissolved7β-(2-thienylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (876 mg) and sodium sulfite (504 mg) and the resultant solution wasstirred for 120 minutes at 60° C. To the reaction mixture was addedwater (10 ml) and the mixture was adjusted its pH to 7.2 with 2 N-HCl,followed by condensation under reduced pressure. The residue wassubjected to column-chromatography of silica-gel and eluted withacetonitrile-water (7:1-5:1). The fractions containing the objectivecompound were combined and the mixture was subjected to distillation ofacetonitrile. The residue was chromatographed on column packed withAmberlite XAD-2 and eluted with water and then water-methanol. Thefractions containing the objective compound were combined and themixture was condensed under reduced pressure, followed by freeze-dryingto give disodium salt of7β-(2-thienylacetamido)-3-sulfomethyl-3-cephem-4-carboxylic acid (310mg).

IR(KBr): 3450, 1760, 1665, 1605, 1190, 1055 cm⁻¹

NMR(δ in D₂ O): 3.67(2H,ABq,J=17 Hz), 3.92(2H,s), 4.16(2H,ABq,J=16 Hz),5.20(1H,d,J=5 Hz), 5.64(1H,d,J=5 Hz), 7.05 & 7.40(3H,m).

EXAMPLE 97

In 50 ml of dichloromethane was dissolved 7.05 g of7-[D-phthalimido-5-carboxyvaleramido]-3-hydroxymethyl-3-cephem-4-carboxylicacid ditriethylamine salt and, at 0° C., triethylamine (1.5 ml) wasadded to the solution, followed by dropping diketene (2.0 ml) during 10minutes at -5°-0° C. The mixture was further stirred for 50 minutes at-5°-0° C. and added 40 ml of water, followed by adjusting its pH to 6.0with 2 N-HCl. The aqueous layer was washed with dichloromethane (10 ml)and 2.25 g of 5-mercapto-1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazole wasadded thereto, followed by adjusting to pH 5.5. The solution was stirredat 60° C. for 40 minutes, and then the insolubles were filtered off. Tothe filtrate were added sodium chloride (15.0 g) and a saturated aqueoussolution of sodium chloride (50 ml) and the mixture was adjusted to pH2.0 with 4 N-HCl. The solid materials precipitated were collected byfiltration and washed with a saturated aqueous solution of sodiumchloride and water in this order, followed by drying to give 6.75 g of7β-(D-5-phthalimido-5-carboxyvaleramido)-3-{1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazol-5-yl}thiomethyl-3-cephem-4-carboxylicacid hydrochloride.

IR(KBr): 3370, 1775, 1715, 1640 cm⁻¹

NMR(δ in d₆ -DMSO): 1.30-2.40(6H,m), 3.5-4.8(9H,m), 5.04(1H,d,J=5 Hz),5.60(1H,q,J=5.8 Hz), 7.90(4H,s), 8.86(1H,d,J=8 Hz)

EXAMPLE 98

In a phosphate buffer solution of pH 6.4 (3 ml) were dissolved5-mercapto-2-methyl-1,3,4-thiadiazole (79 mg), sodium hydrogen carbonate(50 mg) and7β-[2-(1H-tetrazol-1-yl)acetamido]-3-(3-carboxypropionyloxy)methyl-3-cephem-4-carboxylicacid disodium salt (243 mg) and the resultant solution was heated forone hour at 60° C. After cooling, the reaction solution was concentratedunder reduced pressure and the residue was subjected tocolumn-chromatography on Sephadex LH-20(250 ml), elution being carriedout with water. The fractions containing the desired product were pooledand lyophilized. The procedure provided sodium7β-[2-(1H-tetrazol-1-yl)acetamido]-3-(2-methyl-1,3,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylate.

NMR(δ in D₂ O): 2.78(3H,s,--CH₃), 3.60(2H,ABq,J=18 Hz, 2-CH₂),4.25(2H,ABq,J=13 Hz,3-CH₂), 5.12(1H,d,J=4.5 Hz,6-H), 5.58(2H,s,--CH₂CO--), 5.70(1H,d,J=4.5 Hz,7-H), 9.15(1H,s,tetrazole-H)

EXAMPLE 99

In a phosphate buffer of pH 6.4 (3 ml) were dissolved sodium7β-[D-α-t-butoxycarbonylamino-α-(p-hydroxylphenyl)-acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylate(280 mg) and sodium salt of 5-mercapto-1H-1,2,3-triazole (120 mg) andthe resulting solution was heated at 60° C. for 60 minutes. Aftercooling down to room temperature, the reaction solution waschromatographed on a column packed with Amberlite XAD-2 and the combinedeluates containing the desired product was freeze-dried to give sodium7β-[D-α-t-butoxycarbonylamino-α-(p-hydroxyphenyl)acetamido]-3-(1H-1,2,3-triazol-5-yl)thiomethyl-3-cephem-4-carboxylate.

IR(KBr): 3400, 1762, 1678 cm⁻¹

NMR(D₂ O): δ 1.46[9H,s,-C(CH₃)₃ ], 3.00-4.12(4H,m,2-CH₂, 3-CH₂),5.00(1H,d,J=4.5 Hz,6-H), 5.60(1H,d,J=4.5 Hz,7-H), 6.78-7.42(4H,m,##STR110## 7.71(1H,s,triazole-4-H)

Thus obtained sodium7β-[D-α-t-butoxycarbonylamino-α-(p-hydroxyphenyl)acetamido]-3-(1H-1,2,3-triazol-5-yl)-thiomethyl-3-cephem-4-carboxylate(175 mg) was dissolved in formic acid (4 ml) and the resultant solutionwas stirred for 2 hours at room temperature. The reaction solution wassubjected to distillation under reduced pressure and then to azeotropy(3 times) with toluene to remove formic acid, followed by drying overphosphorous pentoxide over night. Thus obtained foamy material wasstirred with water-methanol (8:2)(15 ml) and the mixture was subjectedto filtration, treatment with activated carbon and then filtration usingCelite. The filtrate was freeze-dried to give7-[D-α-amino-α-(p-hydroxyphenyl)acetamido)-3-(1H-1,2,3-triazol-5-yl)thiomethyl-3-cephem-4-carboxylicacid. This product was in good agreement with the standard in thin-layerchromatography and liquid chromatography.

EXAMPLE 100

In water (10 ml) were dissolved7β-[2-(2-imino-4-thiazolin-4-yl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid (908 mg), 5-mercapto-1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazole(450 mg) and sodium hydrogen carbonate (168 mg) and the resultantsolution was heated at 55° C. for 60 minutes. The reaction solution wasrevealed to contain 7β-[2-(2-imino-4-thiazolin-4-yl)acetamido]-3-{1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazol-5-yl}thiomethyl-3-cephem-4-carboxylicacid in 81% yield relative to the starting cephalosporin by liquidchromatography. The reaction solution was adjusted its pH to 5.8 andpurified by column-chromatography using Amberlite XAD-2.

IR(KBr): 1765 cm⁻¹

NMR(D₂ O): δ 3.06(6H,s,-N(CH₃)₂), 3.5-4.8(10H,m), 5.12(1H,d,J=5 Hz,6-H),5.65(1H,d,J=5 Hz,7-H), 6.62(1H,s,thiazolin-H)

EXAMPLE 101

In dimethylformamide (16.0 ml) was suspended monosodium salt monohydrateof deacetylcephalosporin O(4.13 g) and the resultant suspension wasdissolved by adding concentrated hydrochloric acid (1.66 ml) at atemperature lower than 0° C., followed by adding dimethylformamide (16.0ml), triethylamine (4.90 ml) and phthalic anhydride (2.96 g) in thisorder. The resultant mixture was stirred for one hour and a half at 20°C. and the reaction solution was poured onto a mixture of an aqueoussolution of sodium chloride (200 ml) and dichloromethane (40 ml). Themixture was adjusted its pH to 6.5 and the aqueous layer was separated.The layer was washed with dichloromethane and extracted by a mixedsolution of ethyl acetate-tetrahydrofuran (3:1)(50 ml×3) and thecombined extract was washed with a saturated aqueous solution of sodiumchloride, followed by drying over magnesium sulfate. The dried extractwas concentrated under reduced pressure and ether was added to theconcentrate. The procedure provided7β-[D-5-(2-carboxybenzamido)-5-carboxyvaleramido]-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid (6.22 g).

IR(KBr): 1780, 1735, 1725, 1715, 1640 cm⁻¹

NMR(δ in d₆ -DMSO): 1.40-2.40(6H,m), 2.62(2H,ABq,J=18 Hz), 4.35(1H,m),5.09(2H,ABq,J=13 Hz), 5.10(1H,d,J=5 Hz), 5.72(1H,dd,J=5 & 8 Hz),7.30-7.9(8H,m), 8.53(1H,d,J=8 Hz), 8.82(1H,d,J=8 Hz)

What is claimed is:
 1. A compound of the formula ##STR111## wherein R¹is an acyl group and W is acetonyl or --X--OH, in which X is a carbonchain which is capable of forming a five- or six-membered ring with##STR112## which carbon chain may include a double bond or at least oneatom selected from the group consisting of oxygen, nitrogen and sulfur,and on which carbon chain one or more suitable substituents may beattached, or a salt thereof.
 2. A compound as claimed in claim 1,wherein the salt is a pharmaceutically acceptable salt.
 3. A compound asclaimed in claim 2, wherein the pharmaceutically acceptable salt is asalt with an alkali metal, an alkaline earth metal or an amine.
 4. Acompound as claimed in claim 1, wherein the compound is one of theformula ##STR113## wherein R¹ is an acyl group, or a salt thereof.
 5. Acompound as claimed in claim 1, wherein the compound is one of theformula ##STR114## wherein R¹ is an acyl group and X has the samemeaning as defined in claim 42, or a salt thereof.
 6. A compound asclaimed in claim 1, wherein the five- or six-membered ring isO-carboxymandelic anhydride, O-carboxy-α-hydroxypropionic anhydride,O-carboxy-β-hydroxypropionic anhydride, O-carboxy-3-methylsalicylicanhydride, O-carboxy-(α-hydroxy-α-phenyl)propionic anhydride orO-carboxy(α-hydroxy-β-phenyl)propionic anhydride.
 7. A compound asclaimed in claim 1, wherein the five- or six-membered ring issubstituted by carboxyl, halogen, nitro, alkyl having up to 3 carbonatoms, alkenyl having up to 2 carbon atoms, aralkyl, aryl or hydroxy, ormercapto substituted by one of said groups.
 8. A compound according toclaim 1, namely,7β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-mandelyloxymethyl-3-cephem-4-carboxylicacid, or a salt thereof.
 9. A compound according to claim 1, namely,7β-(D-5-phthalimido-5-carboxyvaleramido)-3-mandelyloxymethyl-3-cephem-4-carboxylicacid, or a salt thereof.
 10. A compound according to claim 1, namely,7β-phenylacetamido-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylic acid,or a salt thereof.
 11. A compound according to claim 1, namely,7β-phenoxyacetamido-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 12. A compound according to claim 1, namely,7β-mandelamido-3-mandelyloxymethyl-3-cephem-4-carboxylic acid, or a saltthereof.
 13. A compound according to claim 1, namely,7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 14. A compound according to claim 1, namely,7β-(D-5-benzamido-5-carboxyvaleramido)-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 15. A compound according to claim 1, namely,7β-[D-5-(p-toluenesulfonamido)-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 16. A compound according to claim 1, namely,7β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 17. A compound according to claim 1, namely,7β-(D-5-caprylamido-5-carboxyvaleramido)-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 18. A compound according to claim 1, namely,7β-(2-thienylacetamido)-3-mandelyloxymethyl-3-cephem-4-carboxylic acid,or a salt thereof.
 19. A compound according to claim 1, namely,7β-[2-(2-imino-4-thiazolin-4-yl)acetamido]-3-mandelyloxymethyl-3-cephem-4-carboxylicacid, or a salt thereof.
 20. A compound according to claim 1, namely,7β-(3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 21. A compound according to claim 1, namely,7β-[2-(2-oxo-4-thiazolin-4-yl)acetamido]-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 22. A compound according to claim 1, namely,7β-[D-α-tert-butoxycarbonylamino-α-(p-hydroxyphenyl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid sodium salt.
 23. A compound according to claim 1, namely,7β-(D-α-sulfophenylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a pharmaceutically acceptable salt thereof.
 24. A compoundselected from the group consistingof7β-(2-thienylacetamido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid,7β-(5-amino-5-carboxyvaleramido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid, wherein the amino group may be protected,7β-(5-amino-5-carboxyvaleramido)-3-(3-carboxyacryloyloxy)methyl-3-cephem-4-carboxylicacid, wherein the amino group may be protected, β-(5-amino-5-carboxyvaleramido)-3-(2-carboxy-6 (or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid, wherein the aminogroup may be protected, 7β-(5-amino-5-carboxyvaleramido)-3-(2,4 (or5)-dicarboxybenzoyloxy)methyl-3-cephem-4-carboxylic acid, wherein theamino group may be protected,7β-α-sulfophenylacetamido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid,7β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid,7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid,7β-[D-5-(2-carboxybenzamido)-5-carboxyvaleramido]-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid,7β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-(3-carboxyacryloyloxy)methyl-3-cephem-4-carboxylicacid,7β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid,7β-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid,7β-[D-5-(2-carboxy-6(or3)-nitrobenzamido)-5-carboxyvaleramido]-3-(2-carboxy-6(or3)-nitrobenzoyloxy)methyl-3-cephem-4-carboxylic acid,7β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-(2,4(or5)-dicarboxybenzoyloxy)methyl-3-cephem-4-carboxylic acid, β-[D-5-(p-tert-butylbenzamido)-5-carboxyvaleramido]-3-[3-carboxy-3(or2)-(p-chlorophenylthio)propionyloxy]methyl-3-cephem-4-carboxylic acid,7β-(2-thienylacetamido)-3-(3-carboxyacryloyloxy)methyl-3-cephem-4-carboxylicacid, 7β-(4-chloro-3-oxobutylamido)-3-[2-carboxy-6(or3)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid,7β-phenylacetamido-3-(2-carboxybenzoyloxy)methyl-3-cephem-4-carboxylicacid, and a salt of any one of said compounds.
 25. A compound as claimedin claim 1, wherein the R¹ represents an acyl group of the formula##STR115## wherein R⁴ represents acetyl, halogenoacetyl, phenyl,p-hydroxyphenyl, thienyl, 2-imino-4-thiazolin-4-yl,2-oxo-4-thiazolin-4-yl, tetrazolyl, phenoxy or 3-amino-3-carboxypropyland R⁵ represents hydrogen, sulfo, amino or hydroxy, and each amino andcarboxyl group in R⁴ and R⁵ may be protected.
 26. A compound as claimedin claim 25, wherein the protective group for the amino group isselected from the group consisting of phthaloyl, benzoyl,o-carboxybenzoyl, p-nitrobenzoyl, toluoyl, naphthoyl,p-tert-butylbenzoyl, p-tert-butylbenzenesulfonyl, phenylacetyl,benzenesulfonyl, phenoxyacetyl, toluenesulfonyl, chlorobenzoyl, acetyl,valeryl, capryl, n-decanoyl, acryloyl, pivaloyl, camphorsulfonyl,methanesulfonyl, chloroacetyl, tert-butoxycarbonyl, ethoxycarbonyl,isobornyloxycarbonyl, phenyloxycarbonyl, trichloroethoxycarbonyl,benzyloxycarbonyl, β-methylsulfonylethoxycarbonyl, methylcarbamoyl,phenylcarbamoyl, naphthylcarbamoyl, and 2-methoxycarbonyl-1-methylvinyl.27. A compound according to claim 1, namely,7β-[5-amino-5-carboxyvaleramido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, wherein the amino group may be protected, or a salt thereof.
 28. Acompound according to claim 1, namely,7β-(α-sulfophenylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 29. A compound according to claim 1, namely,7β-(4-chloro-3-oxobutyramido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 30. A compound according to claim 1, namely,7β-(4-bromo-3-oxobutylamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 31. A compound according to claim 1, namely,7β-[2-(2-imino-4-thiazolin-4-yl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 32. A compound according to claim 1, namely,7β-(2-thienylacetamido)-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid or a salt thereof.
 33. A compound according to claim 1, namely,7β-[2-(1H-tetrazol-1-yl)acetamido]-3-(3-oxobutyryloxy)methyl-3-cephem-4-carboxylicacid, or a salt thereof.
 34. A compound according to claim 1, namely,7β-[α-amino-α-(p-hydroxyphenyl)acetamido]-3-(3-oxobutyryloxy)-methyl-3-cephem-4-carboxylicacid, wherein the amino group may be protected, or a salt thereof.